COLUMBIA  LIBRARIES  OFFSITE 

HEALTH  SCIENCES  STANDARD 


HX64140148 
QP535.P1  F74  A  review  of  the  lite 


Cduttibia  Uni  bertfttp 


firfemtr?  Htbrarg 


Digitized  by  the  Internet  Archive 

in  2010  with  funding  from 

Open  Knowledge  Commons  (for  the  Medical  Heritage  Library  project) 


http://www.archive.org/details/reviewofliteratuOOforb 


OHIO  AGRICULTURAL  EXPERIMENT  STATION 


TECHNICAL    SERIES,    BULLETIN    No.    5 


A  REVIEW  OF  THE  LITERATURE 

OF 

PHOSPHORUS  COMPOUNDS  IN  ANIMAL 
METABOLISM 

By  E.  B.  FORBES  AND  M.  HELEN  KEITH 


WOOSTER,  OHIO,  U.  S.  A.,  MARCH,  1914 


OHIO  AGRICULTURAL  EXPERIMENT  STATION 


GOVERNING  BOARD 
THE  AGRICULTURAL  COMMISSION  OF  OHIO 

Columbus 


A.  P.  Sandles,  President 
S.  E.  Strode 


C  G.  Williams 
H.  C.  Price 


B.  P.  Gayman, 

Secretary 


STATION  STAFF 

Charles  E.  Thorne,  M.  S.   A.,  Director 

DEPARTMENTAL  ORGANIZATION 


ADMINISTRATION 

The  Director,  Chiej 
William  H.  Kramer,  Bursar 

D.  W.  Galehouse,  Assistant 
Dora  Ellis,  Mailing  Clerk 
Glenn  Hall,  Engineer 

W.  K.  Greenbank.  Librarian 
W.  J.  Holmes,  Printer 
AGRONOMY 

C.  G.  Williams,    Chiefs 

F.   A.  Welton,  B.  S.,  Associate 

William  Holmes,  Farm  Manager 

C.  A.  Patton,  Assistant 

C.  A.  Gearhart,  B.  S. ,  Assistant 

E.  C.  Morr,   Office  Assistant 

C.  H.  Lebold,  Asst.  Foreman 
ANIMAL  HUSBANDRY 

B.  E.  Carmichael,  M.  S.,   Chief 
J.  W.  Hammond,  M.  S.,  Associate 
Geo.  R.  Eastwood,  B.  S.,  Assistant 
Don  C.  Mote,  M.  S.,  Assistant 

W.   J.  Buss,  Assistant 

Anthony  Russ,  Herdsman 

E.  C.  Schwan,  Shepherd  (Carpenter) 

BOTANY 

A.  D.  Selby,  B.  S.,  Chiej 

True  Houser,  B.  S.,  Assistant  (Germantown) 

P.   K.  Mathis,  Office  Assistant 

D.  C.  Babcock,  A.  B-,  Assistant 
Richard  Walton,  B.  S.,  Assistant 
Alfred  S.  Orcutt,  M.  S.,  Assistant 

CHEMISTRY 

J.  W.  Ames,  M.  S.,  Chief 
Geo.  E.  Boltz,  B.  S.,  Assistant 
J.  A.  Stenius,  B.  S.,  Assistant 
C  J.  Schollenberger,  Assistant 
Miss  Mabel  Corbould,  Assistant 

CLIMATOLOGY 

J.  Warren  Smith,  C/2/V/"(Columbus)2 

C.  A.  Patton,  Observer 

COOPERATION 

The  Director,  Chief 
W   L.  Elser,  B.  S.,  Executive  Assistant^ 
C.  W.  Montgomery,  Assistant 
H.  P.  Miller,  B.  S.,  Comity  Agent  (Ravenna) 
.  P.  L.  Allen-,  A.  B.,  County  Agent  (Burton,) 
W.  M.  Cook,  A.  B.,  County  Agent  (Xenia) 
M.  O.  Bugby,  B.  S.  County  Agent  (Warren) 
A.  L.  Higgins,  B.  S.,  County  Agent  (Das'ton) 
P.  N.  Meeker,  County  Agent  (Hamilton) 
M.  C.  Thomas,   County  Agent  (Troy) 
C.  Ellis  Bundy,  County  Agent  (Paulding) 

DAIRYING 

C.  C.  Hayden,  M.  S.,  Chief 

A.  E.  Perkins,  M.  S.,  Assistant 

T.  R.  Middaugh,  Office  Assistant 

ENTOMOLOGY 

H.  A.Gossard,  M.  S-,  Chief 
J.  S.   Houser,  M.  S.  A.,  Associate 
W.  H.  Goodwin,  M.  S.,  Assistant 
R.  D.  Whitmarsh,  M.  S.  Assistant 
J.  L.  King,  Assistant 


FORESTRY 

Edmund  Secrest,  B.  S.,  Chief 
J.  J.  Crumley,  Ph.  D.,  Assistant 
A.  E.  Taylor,  B.  S.,  Assistant 
J.   W.  Calland,  B.  S.,  Assistant 

D.  E.  Snyder,  Office  Assistant 

HORTICULTURE 

W.  J.  Green,   Vice  Director,   Chief 
F.  H.  Ballou,  Assistant,  (Newark) 

E.  J.  Riggs,  B.  S.,  Assistant,  (Columbus) 
Paul  Thayer,  B.  S.,  Assistant 

C.  W.  Ellenwood,  Office  Assistant 
Ora  Flack,  Foreman  of  Orchards 
W.  E.  Bontrager,  Foreman  of  Grounds 
C.  G.  Laper,  Foreman  of  Greenhouses 

NUTRITION 

E.  B.  Forbes,  Ph.  D.,  Chief 

M.  Helen  Keith,  A.  M.,  Assistant 

F.  M.  Beegle,  B.  S.,  Assistant 
L.  E.  Morgan,  M.  S.,  Assistant 
Charles  M.  Fritz,  B.  S.,  Assistant 


The  Director,  Chief 

C.  G.  Williams,  Associate  in  soil  fertility 

investigations 
J.  W.  Ames,  M.  S.,  Associate  in  soil  chemistry 
George  N.  Coffey,    Ph.   D.,    Associate    in 

charge  of  soil  survey 
E.  R.  Allen,  Ph.  D.,  Associate  in  soil  biology 
H.  Foley  Tuttle,  M.  S.,  Assistant 
A.  Bonazzi,  B.  Agr.,  Assistant 


DIVISION   OF   EXPERIMENT   FARMS 

District  Experiment  Farms 

Northeastern  Test-Farm,  Strongsville. 
Edward  Mohn,  Resident  Manager 

Southwestern   Test-Farm,  Germantown. 
Henry  M.  Wachter,  Resident  Manager 

Southeastern  Test-Farm,  Carpenter. 
H.  D.  Lewis,  Resident  Manager 
Lewis  Schultz,  Horticultural  Foreman 

Northwestern  Test-Farm,  Findlay. 
John  A.  Sutton,  Resident  Manager 

County  Experiment  Farms 

Miami  County  Experiment  Farm.  Troy 
M.  C.  Thomas,  Agent  m  Charge 

Paulding-  County  Experiment  Farm,  Paulding 
C.  Ellis  Bundy,  Agent  in  Charge 
Harry  Ray,  Foreman 

Clermont  County  Experiment  Farm,   Owensville 
Victor  Herron,  Agent  in  Charge 
Howard  Elliott,  Foreman 

Hamilton  County  Experiment  Farm,  Mt.  Healthy 
Victor  Herron,  Agent  in  Charge 


^With  leave  of  absence.    2In  cooperation  with  Weather  Service,  U.  S.  Department  of  Agriculture. 
sIn  cooperation  with  Bureau  of  Plant  Industry,  U.  S.  Department  of  Agriculture. 


PREFACE. 

This  review  of  the  literature  of  phosphorus  metabolism  was 
undertaken  as  a  part  of  a  general  program  of  experimentation  in 
this  field  which  is  being  conducted  by  the  Department  of  Nutrition 
of  this  institution. 

In  our  selection  of  material  we  have  in  mind  the  bearing  of 
this  work  on  practical  human  nutrition  and  animal  husbandry,  and 
have  sought  to  adapt  it  to  the  requirements  of  the  college  graduate 
who  has  an  especial  interest  in  nutrition. 

A  certain  over-emphasis  of  the  subject  of  this  discussion  must 
inevitably  result  from  the  restriction  of  its  scope  to  a  consideration 
of  the  compounds  of  but  a  single  element,  but  we  have  sought 
throughout  the  discussion  to  maintain  correct  perspective  by  indi- 
cating the  connections  of  our  subject  with  those  matters  with  which 
it  stands  in  natural  relationship. 

Since  the  compounds  of  phosphorus  have  to  do,  in  prominent 
ways,  with  all  of  the  life  processes  of  animals,  our  subject,  though 
narrow  in  form,  might  easily  be  construed  to  cover  the  whole  field 
of  nutrition.  It  has  been  necessary,  therefore,  in  the  conduct  of 
this  study,  arbitrarily  to  limit  its  scope  in  order  to  render  its  com- 
pletion in  any  way  a  practical  possibility,  and  we  have  been  obliged 
entirely  to  ignore  some  important  divisions  of  the  subject  for  no 
better  reason  than  that  life  is  short. 

The  thoroughness  with  which  we  have  treated  the  several  sec- 
tions of  this  discussion  varies  much.  Our  treatment  of  the  field 
of  pathology  has  been  especially  incomplete.  Here  it  was  our  idea 
to  include  only  such  matters  as  we  considered  of  interest  to  the 
student  of  nutrition  in  general,  and  we  have  not  gone  into  a  discus- 
sion of  diseases,  as  such,  in  a  way  to  serve  the  requirements  of  the 
pathologist.  We  have  made  no  careful  search  of  the  literature  of 
pathology  for  materials  on  phosphorus  metabolism.  The  whole 
subject  of  phosphorus  poisoning  has  been  omitted,  this  matter  being 
considered  as  quite  apart,  in  reality,  from  phosphorus  metabolism  in 
the  usual  sense — and  the  literature  is  enormous.  The  relation  of 
phosphatids,  especially  lecithin,  to  the  action  of  alkaloids  and  other 
drugs,  as  well  as  antitoxins,  precipitins,  hemolysins,  bacteriolysins, 
agglutinins  and  opsonins  is  of  a  special  nature,  having  to  do  with  the 
maintenance  of  .immunity  rather  than  with  phosphorus  metabolism 
in  the  usual  sense,  and  is  not  included  in  this  discussion. 


4  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Clinical  reports  are  submitted  in  great  brevity  and  usually 
without  comment.  They  certainly  have  a  value,  but  it  is  impos- 
sible to  say,  in  a  given  case,  whether  this  value  be  great  or  small. 
Some  of  the  reasons  for  this  are  as  follows : 

The  first  interest  of  the  medical  profession  is  the  cure  of  the 
patient  and  not  the  acquirement  of  experimental  evidence ;  it  is 
usually  impossible  to  provide  proper  experimental  controls  or 
checks;  even  in  hospitals  the  environment  of  the  patient  does  not 
always  constitute  a  proper  basis  for  exact  experimentation;  the 
rights  of  the  patient  prevent  the  full  development  and  realization  of 
such  unfavorable  results  of  treatment  as  might  follow  under  strict- 
ly experimental  conditions;  and  the  imagination  of  the  patient  un- 
doubtedly cuts  a  figure  in  symptoms  and  results  which  is  not  char- 
acteristic of  experiments  on  animals  with  less  capacity  to  think. 

The  chemistry  of  phosphorus  compounds,  and  the  phosphorus 
compounds  of  animal  bodies  and  products,  and  of  foods  and  drugs 
have  been  briefly  considered  with  the  idea  of  making  clear  the  bear- 
ing of  the  matter  on  normal  phosphorus  metabolism,  in  which  field 
we  have  considered  all  of  the  material  which  has  come  to  our  atten- 
tion. 

The  selection  of  articles  for  inclusion  in  this  review  was  made 
by  the  use  of  indices,  supplemented  by  a  page-to-page  search  of  the 
journals  considered  as  most  likely  to  contain  the  desired  material. 
The  main  sources  of  the  articles  included  are  as  indicated  in  the  fol- 
lowing list,  though  many  references  were  also  taken  from  scattered 
sources. 

American  Journal  of  Physiology  1  (1898)  to  32,  (Dec.  1913) 

Archiv  fur  die  gesammte  Physiologie  1  (1868)  to  156,  442  (Mar.  1914) 

Biochemical  Journal  1  (1906)  to  7,  (Dec.  1913) 

Biochemische  Zeitschrift  1  (1906)  to  59  (Feb.  1914) 

Chemical  Abstracts  1  (1907)  to  8,  1220  (Mar.  20,  1914) 

Die  Ernahrung  der  landwirtschaftlichen  Nutztiere,  vierte  Auflage;  O.  Ilell- 

ner  1907. 

Ergebnisse  der  Physiologie  1  to  13  (1913) 

Experiment  Station  Record  (1899)  to  29,  899  (Feb.  1914) 

Handbuch  der  Biochemie  des  Menschen  unci  der  Tiere,  Carl  Oppenheimer, 

1908-1913. 

International  Catalog  of  Scientific  Literature.       Q.       Physiology  1  to  8 

(1911) 

Jahresbericht  fiber  die  Fortschritte  der  Thierchemie  1  (1871)  to  41  (1911) 
Journal  fur  Landwirtschaft  49  (1901)  to  61  (1913) 

Journal  of  the  American  Medical  Association  34  (1900)  to  61  (Dec,  1913) 
Journal  of  Physiology  26  (Dec.  1900)  to  47,  No.  6  (Feb.  1914) 
Metabolism  and  Practical  Medicine,  Vols.  I,  II,  III,  Carl  von  Noorden,  1907. 
Physiologie  und  Pathologie  des  Mineralstoffwechsels,  Albert  Albu  and  Carl 

Neuberg,  1906. 

Review  of  American  Chemical  Research  6  (1900)  to  12  (1906)— last  issue. 
Revue  de  la  Societe  d'  Hygiene  Alimentaire  1  (1904)  to  7  (1909)  entire. 
The  Journal  of  Biological  Chemistry  1  (1905-06)  to  17,  304  (Mar.  1914) 


PHOSPHORUS  METABOLISM  5 

The  Journal  Of  the  American  Chemical  Society  1  (1879)  to  36,  616  (Mar. 

1914) 

Vierteljahreschrift  der  Nahrungs- Chemie  9  (1894)  to  12  (1897)  last  issue. 
Zeitschrift  fur  Biologie  39  (N.  F.  21)  to  63  (N.  F.  45),  Heft  9  (Mar.  1914) 
Zeitschrift  fur  Kinderheilkunde.       Originate  1   (1910-11)  to  10,  Hefte  1-4 

(Feb.  1914) 

Zeitschrift  fur  Nahrungs  Untersuchung  1   (1898)  to  10   (1905) 
Zeitschrift  fur  physiologische  Chemie  1  (1877)  to  89,  324  (Feb.  1914) 
Zeitschrift  fur  Untersuchung  der  Nahrungs-  und  Genussmittel  to  27,  360 

(Feb.  1914) 

In  general  we  have  considered  only  original  articles.  Most  of 
the  study  of  the  literature  has  been  pursued  by  a  uniform  method 
of  preparation  of  abstracts  including  (1)  the  complete  reference, 
(2)  the  object  of  the  investigation,  (3)  the  methods  employed,  (4) 
the  nature  of  the  data  recorded,  (5)  numerical  results  and  (6)  the 
author's  conclusions  in  full. 

Gross  misconceptions  have  been  generally  ignored,  in  consider- 
ation of  the  extreme  brevity  with  which  we  have  been  obliged  to 
treat  much  good  material. 

We  have  not  gone  into  a  discussion  of  analytical  methods, 
though  they  have  been  considered,  and  much  work  has  been  thrown 
out  because  of  the  use  of  methods  unsuited  to  the  purpose.  It  is 
true,  however,  that  much  work,  known  to  have  been  accomplished 
by  faulty  methods,  has  been  retained,  often  because  the  methods 
have  been  fairly  satisfactory  for  comparative  purposes,  and  again, 
frequently  because  better  methods  have  not  been  discovered  or  have 
not  come  into  general  use.  In  very  few  cases,  for  instance,  do  the 
differential  estimations  of  the  groups  of  organic  phosphorus  com- 
pounds rest  upon  critically  established  analytical  procedures,'  and 
even  so  simple  a  problem  as  the  determination  of  inorganic  phos- 
phorus in  plant  and  animal  substances  is  still  a  problem,-  fairly  well 
settled  for  some  substances,  it  is  true,  but  by  no  means  for  all  from 
which  we  report  determinations. 

Candor  requires  the  observation  that  much  of  the  work  con- 
sidered is  without  great  value,  but  until  supplanted  by  results  of 
higher  grade  we  are  obliged  to  give  it  space.  This  applies  espec- 
ially to  metabolism  data,  the  experiments  being,  as  a  rule,  much  too 
short  to  afford  safe  bases  for  conclusions ;  and  elaborate  discussions 
of  urinary  conditions,  feces  data  not  being  included,  are  far  too 
common. 

,  Numerical  data  have  been  largely  retabulated,  and  very  com- 
monly they  have  been  recalculated.  If  found  inconsistent  they 
have  usually  been  thrown  out,  but  have  sometimes  been  corrected, 
with  the  author's- approval. 


6  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

In  addition  to  data  on  phosphorus  metabolism,  we  have  also 
commonly  taken  from  the  general  studies  the  data  on  calcium  and 
nitrogen,  since  these  elements  are  more  prominently  involved  than 
others  with  the  metabolism  of  compounds  of  phosphorus. 

We  are  glad  to  acknowledge  here  the  assistance  in  abstracting 
received  from  B.  M.  Hendrix,  Dr.  H.  W.  Houghton,  E.  C.  Lathrop, 
Grace  Mac  Leod,  Dr.  Martha  A.  Phelps,  Dr.  J.  B.  Rieger,  Florence  C. 
Sargent,  Marion  E.  Sparks  and  Dr.  G.  M.  Tucker. 

The  abstracting  has  been  done  in  the  following  institutions : 
Ohio  Agricultural  Experiment  Station,  University  of  Illinois,  Li- 
brary of  Congress,  Surgeon  General's  Library,  Library  of  the  De- 
partment of  Agriculture,  New  York  Academy  of  Medicine,  New 
York  Public  Library,  Columbia  University,  Massachusetts  Institute 
of  Technology,  Boston  Medical  Library,  Harvard  Medical  Library,- 
Boston  Public  Library,  Harvard  University,  Boston  Natural  His- 
tory Museum,  Connecticut  Agricultural  Experiment  Station,  Yale 
University,  Cleveland  Medical  Library  and  Western  Reserve  Med- 
ical College. 


PHOSPHORUS  METABOLISM 


TABLE  OF  CONTENTS 


Part  I    Chemistry  of  Organic  /Compounds  of  Phosphorus  Page 

1 .  Nucleoproteins  and  Their  Derivatives 13 

2 .  Phosphoproteins     32 

3 .  Phosphocarnic  Acid   49 

4.  Phytin     51 

5 .  Phosphatids  58 

Part  II    The  Phosphorus  of  Foods 

1 .  General  Discussion   76 

2 .  Organic  and  Inorganic  Phosphorus  in  Foods   80 

3 .  Phytin  in  Foods 81 

4 .  Lecithins  in  Foods 84 

5 .  Nuclein  Phosphorus   in   Foods    90 

6.  Pyrophosphoric  Acid  in  Foods  91 

7 .  Proprietary  Preparations    92 

8 .  Effects  of  Water  on  the  Phosphorus  of  Forage  Plants 93 

9.  Effects  of  Fertilizers  on  the  Composition  of  Foods 95 

10 .  Addition  of  Phosphates  to  Silage 101 

11.  References  to   Extensive  Presentations   of   Total   Phosphorus 

Determinations  in  Foods  ...,.: 103 

Part  III     The  Phosphorus  of  Animal  Bodies  and  Products 

1 .  General  Studies,  Grown  Men  and  Animals 104 

2 .  Fetuses  and  New-Born  Young   108 

3 .  Bones,  Teeth,  Marrow,  Cartilage  Ill 

4.  Muscle    : 120 

5.  Feathers 126 

6.  Brain,  Nerves,  Cerebrospinal  Fluid   127 

7.  Liver  136 

8.  Spleen 139 

9 .  Pancreas 140 

10.  .   Kidney    141 

11 .  Suprarenal  Capsules   143 

12.  Thymus     143 

13 .  Thyroid 144 

14 .  Lungs    145 

15 .  Hypophysis    146 

16 .  Digestive  Mucosa    146 

17.  Genitalia     146 

18.  Blood     149 

19.  Milk     153 

20.  Eggs    169 

21 .  Digestive  Secretions   172 

22.  Chyle     177 

23.  Lymph    177 


8  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Part  rV    Normal  Phosphorus  Metabolism  Page 

1 .  Maintenance  of  Neutrality   178 

2.  The  Absorption  and  Elimination  of  Compounds  of  Phosphorus 

General  Consideration    181 

The  Influence  of  Phosphates  on  Digestion 186 

Autolysis  of  Compounds  of  Phosphorus   187 

Method  of  Urinary  Elimination  of  Phosphates  188 

Organic  Phosphorus  in  the  Urine  190 

Phosphorus  Compounds  in  the  Feces 194 

3.  Phosphorus  Excretion  as  Affected  by  Various  Conditions 

Acids  and  Acid  Salts    198 

Sodium  and  Potassium 200 

Calcium  and  Magnesium  203 

Diet  and  Species 207 

Summary   211 

4.  Relative  Metabolism  of  Phosphorus  and  Other  Elements 213 

5.  Metabolism  Experiments  With  Inorganic  Phosphates  221 

6.  Metabolism  of  Nucleoproteins  and  Nucleic  Acids 229 

General  Discussion   , 229 

Digestion  Studies  with  Enzymes  from  the  Alimentary 

Tract     231 

Effects  of  Bacteria  in  the  Gastrointestinal  Tract 234 

Ferments  of  Individual  Organs 235 

Animal  Experiments  on  the  Digestion  of  Nucleoproteins 

and  Nucleic  Acids  237 

Accompaniments  of  the  Ingestion  or  Injection  of  Nuclear 

Material     245 

Significance  of  Nucleoproteins  and  Nucleic  Acids  as  Active 

Agents  in  Specific  Body  Processes 255 

Therapeutic  and  Prophylactic  Use  of  Nucleins  and  Nucleic 

Acids ; 256 

Summary   258 

7.  Metabolism  of  Casein 

Production  of  Casein  in  Milk  Glands   260 

Digestion  of  Casein 261 

Animal  Experiments  With  Casein   272 

Summary 274 

8.  Metabolism  of  the  Compounds  of  Glycerophosphoric  Acid 

The  Function  of  Lecithin  in  Intermediary  Metabolism. . .  .276 

The  Influence  of  Lecithin  in  Digestion 282 

Summary   286 

The  Digestion  of  Lecithin  and  Glycerophosphates 286 

Summary 289 

Balance  Experiments  With  Lecithin  and  Other  Compounds 

of  Glycerophosphoric  Acid   290 

Summary   295 

Growth  and  Composition  of  Animals  as  Affected  by  Com- 
pounds of  Glycerophosphoric  Acid 296 

Summary   303 

9.  Metabolism  of  Phosphocarnic  Acid 303 


PHOSPHORUS  METABOLISM  9 

Page 

10.  Metabolism  of  Phytin 

Feeding  Experiments  with  Men  and  Animals 305 

Clinical  Experiments  with  Phytin 315 

Summary   317 

11.  The  Nutritive  Values  of  Organic  and  Inorganic  Phosphorus 

Metabolism  Experiments  with  Men  and  Animals 318 

Miscellaneous  Experiments  , 318 

Organic  Phosphorus  Synthesis  in  Fasting  Salmon  . . .  .354 
Advantage   of   Complexity  of   Organization   in   Food 

Phosphorus    . 355 

Feeding  Experiments  with  Rats  and  Mice  on  Rations 

Composed  of  Simple  Purified  Nutrients 357 

Summary   364 

12.  Common  Foods  in  Relation  to  Phosphorus  Metabolism 

Miscellaneous  Experiments  with  Men  and  Animals 366 

The  Phosphorus  Content  of  Milk  as  Affected  by  Foods ....  379 
Diet  and  the  Phosphates  of  the  Bones 384 

13.  Phosphorus  Requirements  of  Animals 

General  Discussion   397 

Phosphorus  Excretion  During  Fast   399 

Normal  Phosphorus  Requirements  of  Adult  Human 

Beings     403 

Phosphorus  Requirements  of  Infants    414 

"  Cattle 419 

"  Horses   422 

"  Swine 424 

"  Sheep   425 

"   Dogs    426 

"  Rats  428 

14.  Phosphorus  Metabolism  as  Affected  by  Various  Conditions 

Altitude    429 

Amount  of  Food   430 

Fast .438 

Incubation  446 

Infancy  449 

Mental  Work    461 

Nerve  Stimulation    463 

Physical  Exercise    464 

Pregnancy   470 

Sleep     472 

Thirst  and  Water  Drinking 473 

Time  of  Day   474 

Part  V    Phosphorus  Metabolism  in  Disease 

1 .  Acid   Intoxication    476 

2.  Acromegaly     479 

3 .  Alcoholism     483 

4.  Arthritis     484 

5.  Beriberi 484 


10  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Page 

6.  Blood  in  Relation  to  Pathological  Phosphorus  Metabolism 

Chlorosis  and  Anaemia 492 

Leukaemia     493 

Blood   Dissolution    496 

Phosphorus  of  the  Blood  as  Affected  by 

Anaemia     496 

Chlorosis    496 

Leukaemia     497 

Various  Diseases   497 

Nuclein  Therapy  in  Anaemia 502 

7.  Calcareous  Degeneration  503 

8.  Cancer    503 

9 .  Caries    504 

10 .  Castration  and  Ovariotomy 505 

11.  Composition  of  the  Body  in  Disease 506 

12.  Diabetes   Mellitus    510 

13 .  Endoarteriitis    512 

14 .  Fatty  Degeneration 512 

15.  Fever    ! 514 

16 .  Glycerophosphate  Therapy   520 

17.  Gout    522 

18 .  Hemorrhage    526 

19 .  Hookworm   527 

20.  Icterus    527 

21 .  Lecithin  Therapy 528 

22 .  Malnutrition  of  the  Bones   534 

23.  Mental  and  Other  Nervous  Disorders 

Nerve  Degeneration   536 

Epilepsy   540 

Paralysis    541 

Insanity    543 

Administration  of  Phosphorus  Compounds  and  Other 

Drugs  in  Mental  and  Other  Nervous  Diseases 545 

24.  Nephritis 547 

25.  Nuclein  Therapy   549 

26.  Oedema     549 

27.  Organic  Phosphorus-Containing  Compounds  and  Preparations, 

Effects  on  Phosphorus  Metabolism    549 

28 .  Osteomalacia     552 

29 .  Parathyroidectomy    559 

30.  Phosphaturia     561 

31 .  Phytin  Therapy   566 

32.  Rachitis    566 

33 .  Thyroid  Glands  in  Relation  to  Phosphorus  Metabolism 577 

34 .  Toxins  and  Antitoxins   582 

35 .  Tuberculosis    583 


INTRODUCTION 

As  agricultural  scientists  our  interest  in  the  mineral  elements 
lies  in  that  larger  intermediary  metabolism  between  the  soil  and  the 
sea  which  begins  with  the  weathering  of  the  rocks,  includes  the 
whole  of  plant  and  animal  metabolism,  and  ends  with  the  formation 
of  new  rocks. 

Throughout  this  vast  sweep  of  chemical  change  the  mineral 
elements  occupy  a  unique  and  dominating  position,  entering  in  es- 
sential ways  into  every  process,  and  exerting  an  influence  in  metab- 
olism entirely  out  of  proportion  to  the  amounts  in  which  they  are 
involved. 

In  a  large  and  general  way  life  may  be  regarded  as  a  coordin- 
ated system  of  responses  to  electrical  stimulation.  The  ions,  and  es- 
pecially the  inorganic  ions,  are  the  bearers  of  this  electricity,  and 
it  is  because  of  this  fact  that  they  are  able  to  play  a  leading  role  in 
the  direction  of  the  whole  process  of  metabolism. 

Among  the  several  inorganic  elements  involved  in  animal  life 
phosphorus  is  of  especial  interest.  No  other  one  enters  into  such  a 
diversity  of  compounds  and  plays  an  important  part  in  so  many 
functions.  Structurally,  it  is  important  as  a  constituent  of  every 
cell  nucleus  and  so  of  all  cellular  structures ;  it  is  also  prominent  in 
the  skeleton,  in  milk,  in  sexual  elements,  glandular  tissue  and  the 
nervous  system.  Functionally,  it  is  involved  in  all  cell  multiplica- 
tion, in  the  activation  and  control  of  enzyme  actions,  in  the  main- 
tenance of  neutrality  in  the  organism,  in  the  conduct  of  nerve  stim- 
uli, and  through  its  relation  to  osmotic  pressure,  surface  tension 
and  imbibation  of  water  by  colloids  it  has  to  do  with  the  movement 
of  liquids,  with  the  maintenance  of  proper  liquid  contents  of  the 
tissues,  with  cell  movements  and  with  absorption  and  secretion. 

Throughout  the  intricacies  of  these  processes — in  considering 
the  relations  of  the  animal  to  its  food — let  it  be  our  point  of  view 
that  inheritance  has  furnished  the  plans,  the  details  and  specifica- 
tions which  are  to  govern  the  whole  course  of  metabolism;  that 
food  builds  the  structure  and  maintains  its  processes,  in  so  far  as 
made  possible  by  the  nature  and  amounts  of  its  constituents ;  that 
variability  in  the  composition  and  functions  of  the  animal  body,  and 
excess  of  capacity  in  its  structures  constitute  a  provision  of  safety, 
a  means  of  adaptive  response  to  changes  in  dietary  conditions ;  that 
time  lends  to  these  adaptations  such  permanency,  in  the  individual, 
as  to  constitute  specific  effects  of  foods  on  the  life  and  structure  of 


12  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

the  animal ;  that  these  specific  effects  of  foods  are,  in  general,  due 
rather  to  their  limitations  than  to  stimulation  of  .supernormal  func- 
tion ;  that  the  nature  and  possible  extent  of  these  effects  have  been 
separately  determined  for  each  species  by  the  particular  conditions, 
and  the  variability  of  conditions  of  life  to  which,  through  the  ages, 
they  have  become  adapted,  and  that  in  relation  to  practical  animal 
nutrition  our  interests  are  in  the  highest  states  of  function  rather 
than  in  irreducible  physiological  minima,  since  the  whole  range  of 
success  and  profit  lies  close,  and  ever  closer,  to  maximum  possibili- 
ties. 

The  following  discussion  attempts  no  complete  picture  or  con- 
tinuous account  of  phosphorus  metabolism,  but  seeks  merely  to  ar- 
range in  natural  and  useful  sequence  the  fragmentary  material  of 
which  it  is  composed. 


BULLETIN 

OF  THE 

Ohio  Agricultural  Experiment  Station 

Number  5  March,  1914 

PART  I 

CHEMISTRY  OF  ORGANIC  COMPOUNDS 
OF  PHOSPHORUS 

NUCLEOPROTEINS  AND  THEIR  DERIVATIVES 

Place  in  the  Scheme  of  Chemical  Compounds.  The  facts  as  to 
the  method  of  growth  of  the  individual  animal  lead  us  almost 
certainly  to  the  conclusion  that  the  transmission  of  characters  from 
generation  to  generation,  and  the  determination  of  the  nature  of  all 
cellular  growth  has  as  its  material  basis  the  chemical  structure  of 
the  nucleoproteins  of  the  cell.  In  comparison  with  the  almost 
inconceivable  complexity  of  composition  implied  by  this  idea  our 
knowledge  of  the  chemistry  of  the  nucleoproteins  is  especially 
superficial  and  our  methods  of  study  most  violent.  This  field  must 
remain  for  all  time  in  large  part  unknown  and  unknowable,  and  it 
is  only  with  the  grossest  facts  as  to  composition,  as  made  manifest 
by  the  destruction-of  the  compounds  and  the  study  of  the  wreckage 
products  that  we  are  here  concerned. 

Nucleoproteins  are  those  compound  proteins  which  yield  protein 
and  nucleic  acid  on  cleavage.  Any  of  the  simple  proteins  may  occur 
in  this  union;  and  the  number  of  nucleoproteins  is  practically 
unlimited.  Of  those  which  are  found  in  natural  products  the 
protein  has  been  identified  in  but  very  few  cases. 

Under  the  action  of  weak  acids  or  of  gastric  juice  the  cleavage 
of  nucleoproteins  does  not,  in  most  cases,  produce  a  nucleic  acid 
directly ;  but  instead,  a  protein  compound  of  nucleic  acid  containing 
less  of  protein  and,  consequently,  a  larger  proportion  of  phosphorus. 
It  is  customary  to  call  these  simpler  protein  compounds  of  nucleic 
acids  by  the  name  nucleins,  and  to  let  the  name  nucleoprotein  apply 
to  the  naturally  occurring  substances  which  are  combinations  of 
nucleins  with  further  protein.  Nucleoprotein  as  defined  above 
includes  both  of  these  groups. 

(13) 


14  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

The  nucleins  are  decomposed  by  caustic  alkali  into  protein  and 
nucleic  acids.  All  nucleic  acids  are  rich  in  phosphorus,  and  on 
cleavage  yield  phosphoric  acid,  purin  bases,  a  carbohydrate  (or  car- 
bohydrate derivative)  and  usually  pyrimidin  bases. 

The  paranucleins,  or  pseudonucleins,  are  derived  from  the  phos- 
phoproteins  (nucleoalbumins)  in  the  same  way  that  nucleins  are  de- 
rived from  nucleoproteins,  and  in  certain  cases  paranucleic  acids 
have  been  obtained,  corresponding  to  nucleic  acids.  A.  Kossel 
(1891a),  proposed  the  name  paranuclein,  and  noted  that  para- 
nucleins differ  from  true  nucleins  in  not  yielding  purin.  Giertz 
(1899)  says  that  they  may  be  definitely  distinguished  from  true 
nucleins  in  that  they  readily  dissolve  in  water  made  slightly  alkaline 
with  baryta,  and  that  they  are  decomposed  by  excess  of  baryta  with 
the  splitting  off  of  phosphoric  acid. 

Some  have  reported  the  cleavage  of  nucleoprotein  directly,  with 
the  separation  of  nucleic  acid,  and  either  protein,  or  its  peptones 
and  amino  acids  (Altmann,  1889;  A.  Kossel,  1893;  Umber,  1901). 

The  reader  is  referred  to  the  following  articles  for  reviews  of  the  work  on 
this  subject  reported  up  to  the  dates  of  the  several  papers: — Nolf  (1898), 
Steudel  (1907a),  Schaumann  (1910),  Brugsch  and  Schittenhelm  (1910)  and 
Brahm  (1913). 

Occurrence  of  Nucleoproteins .  Nucleoproteins  occur  chiefly  in 
the  cell  nuclei ;  but  they  also  often  occur  in  the  cytoplasm,  and  are 
found  in  the  blood  serum  and  other  fluids. 

They  have  been  obtained  most  abundantly  from  those  products 
and  organs  which  are  rich  in  nuclei.  The  nuclein  first  discovered 
was  that  formed  from  pus  by  peptic  digestion.  This  discovery  was 
the  work  of  Miescher.  Plosz  (1871  and  Ibid.,  p.  441),  also  obtained 
a  similar  substance  from  the  blood  corpuscles  of  a  bird  and  of  a 
snake.  Miescher's  more  extensive  study,  to  which  we  owe  consid- 
erable of  our  knowledge  of  these  compounds,  was  made  upon  sperm 
and  spermatozoa,  chiefly  from  salmon.  The  substance  which  Mies- 
cher (1878,  1896,  1897)  called  protein-free  nuclein  was  named 
nucleic  acid  by  Altmann  (1899)  on  account  of  its  acid  properties. 

The  nucleic  acids  to  which  the  greatest  amount  of  attention 
has  been  given  are  those  of  the  thymus,  of  spermatozoa  and  of 
yeast ;  and  in  addition  to  these  we  are  fairly  familiar  with  those  of 
the  pancreas,  adrenals,  spleen,  testes  and  fish  eggs.  The  only  plant 
nucleic  acid  besides  that  present  in  yeast,  which  has  been  much  in- 
vestigated, is  that  which  T.  B.  Osborne  and  his  associates  have  iso- 
lated from  wheat  embryo,  and  have  named  tritico-nucleic  acid  (Os- 
borne and  Campbell,  1900a ;  Osborne  and  Harris,  1902 ;  Wheeler  and 
Johnson,  1903;  Mendel,  Underhill  and  White,  1903;  Osborne  and 
Heyl,  1908;  Levene  and  LaForge,  1910). 


PHOSPHORUS  METABOLISM  15 

Other  early  discussions  and  reports  of  nucleins  besides  those 
of  Miescher  and  of  Plosz  are  those  of  J.  W.  Miiller  (1873),  of  Piccard 
(1874)  on  salmon  sperm,  of  von  Jaksch  (1876)  on  nucleins  from 
human  brain,  and  of  Klinkenberg  (1882)  on  some  nucleins  from 
various  seeds  used  as  fodders.  As  long  ago  as  1847  Justus  Liebig 
isolated  from  flesh  an  acid  to  which  he  gave  the  name  inosinic  acid, 
and  which  he  analyzed,  but  he  did  not  find  the  phosphorus.  In 
1878  Schiitzenberger  (1878)  named  decomposition  products  of 
brewer's  yeast,  which  were  such  as  are  now  recognized  as  those  of 
nucleoproteins. 

PROPERTIES  OF  NUCLEOPROTEINS 

The  nucleoproteins,  after  isolation  and  purification,  are  loose, 
white  powders,  insoluble  in  water,  but  soluble  in  alkali  solutions  by 
union  with  the  alkali.  From  such  solutions  they  are  precipitated 
by  acids ;  excess  of  mineral  acids  dissolves  them,  but  acetic  acid  less 
readily.      They  are  acid  in  character. 

The  percentage  composition  of  nucleoproteins  is  very  variable 
and  the  determinations  doubtful;  but  all  contain  phosphorus,  per- 
haps from  0.5  to  3.0  percent.  That  obtained  by  Liebermeister 
(1906)  from  the  blood  serum  of  the  horse  was  very  low  in  phos- 
phorus, only  about  0.08  percent. 

It  has  been  commonly  stated  that  the  nucleoproteins  contain 
iron  in  organic  combination;  but  the  amounts  found  vary  greatly, 
and  the  nature  of  its  relation  to  the  molecule,  if  it  be  a  part  of  the 
nucleoprotein  molecule,  is  not  known  (see  S.  S.  Zaleski,  1886 ;  Ham- 
marsten,  1894 ;  Scaff idi,  1908 ;  Salkowski,  1909a,  1909b ;  Capezzuoli, 
1909a).  Salkowski  (1909b)  finds  iron  present,  but  very  loosely 
bound.  Ascoli  (1899)  reports  it  in  plasminic  acid,  showing  rela- 
tions which  he  thinks  indicate  a  metaphosphoric  combination;  but 
Sauerland  (1910)  fails  to  find  iron  in  the  free  nucleic  acids  from 
pancreas  and  spermatozoa  heads,  and  doubts  its  existence  in  the 
nucleoproteins. 

The  cleavage  products  are  characteristic  constituents.  The 
only  attempts  which  we  have  noticed,  to  make  complete  determina- 
tions of  all  cleavage  products  are  those  of  Wohlgemuth  (1903, 
1904a,  1904b,  1905).  The  nucleoproteins  give  the  color  reactions 
of  proteins.  So  far  as  examined,  they  are  all  dextrorotatory  (Gam- 
gee  and  Jones,  1903a,  1903b,  1903c). 

On  peptic  digestion  nucleoproteins  yield  true  nucleins.  Weak 
acids  cause  decomposition,  and  the  solutions  of  the  alkali  combina- 
tions are  decomposed  even  by  heating  with  water. 


16  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

A  list  of  references  to  nucleoprotein  investigations  is  given 
below : 

NUCLEOPROTEIN  STUDIES  ARRANGED  WITH  REFERENCE  TO  THE 
SOURCE  OF  THE  NUCLEOPROTEIN 

From  Thymus:  Bang,  1900a,  1900b,  1903  and  1904;  Gamgee  and  Jones; 
1903a,  1903b,  1903c;  Halliburton,  1895;  Halliburton  and  Brodie,  1894;  Huis- 
kamp,  1901a,  1901b,  1903;  A.  Kossel,  1900;  Lilienfeld,  1894;  Malengreau,  1900, 
1901;  Steudel,  1913b. 

From  Liver:  Goubau,  1911;  Halliburton,  1892,  1895;  Salkowski,  1909a, 
1909b;   Scaffidi,  1908,  1909a;  Wohlgemuth,  1903,  1904a,  1904b,  1905. 

From  Pancreas:  Gamgee  and  Jones,  1903a,  1903b,  1903c;  Hammarsten, 
1893,  1894;  Jones  and  Whipple,  1902;  Knopf,  1914;  Levene,  1904;  Levene  and 
Stookey,  1904;  Umber,  1900,  1901. 

From  Suprarenals:  Gamgee  and  Jones,  1903a,  1903b,  1903c;  Jones  and 
Whipple,  1902. 

From  Brain:     Halliburton,  1895;  von  Jaksch,  1876;  Levene,  1899. 

From  Blood  Corpuscles:  Bang,  1903,  1904;  Halliburton,  1895;  Halliburton 
and  Friend,  1889;  A.  Kossel,  1881,  1882;  Plosz,  1871. 

From  Blood  Serum:     Liebermeister,  1906;  Pekelharing,  1895. 

From  Leucocytes:    Bang,  1903,  1904;  Lilienfeld,  1894. 

From  Yeast:     A.  Kossel,  1879,  1880,  1881. 

From  Other  Sources:  Spleen;  Capezzuoli,  1909a;  Goubau,  1911;  Sato,  1909. 
Thyroid;  Oswald,  Ad.,  1899.  Kidney;  Goubau,  1911;  Halliburton,  1892,  1895. 
Mammary  glands;  R.  Odenius,  1899.  Human  Placenta;  Bottazzi,  1903;  Cocchi, 
1901.  Testes;  Goubau,  1911.  Spermatozoa;  Steudel,  1911a,  1911b,  1913a. 
Lymph  glands;  Bang,  1903,  1904.  Muscle;  Pekelharing,  1896.  Marrow;  Halli- 
burton, 1895.  Pus;  Goubau,  1911;  A.  Kossel,  1881.  Egg- yolk;  A.  Kossel, 
1885,  1886.  Several  nucleins;  Klinkenberg,  1882.  Barley  sprouts;  Petit, 
1893.       Several  seeds;  Vorbrodt,  1910. 

PROPERTIES  OF  NUCLEIC  ACIDS 

Nucleic  acids  are  white,  amorphous,  acid  powders,  insoluble  in 
water,  but  soluble  in  ammoniacal  or  alkaline  water.  Since  all  of 
the  phosphorus  of  the  nucleoprotein  molecule  is  in  the  nucleic  acid 
fraction,  the  percentage  here  is  high,  perhaps  8.0-10.0  percent. 
The  nucleic  acids  precipitate  proteins  from  solution.  The  affinity 
for  protein  is  perhaps  their  most  significant  property. 

Nucleins  stand  between  nucleoproteins  and  nucleic  acids  in 
properties  and  composition,  corresponding  with  their  intermediate 
position  in  production. 

Feulgen  (1912,  1913a,)  has  prepared,  analyzed  and  studied  the 
properties  of  compounds  of  nucleic  acid  with  dyes,  the  composition 
of  which  he  thinks  is  such  as  to  indicate  the  formation  of  tetrabasic 
salts,  and  he  suggests  that  perhaps  nucleoproteins  are  such  saltlike 
combinations  of  nucleic  acids  with  the  basic  proteins.  Feulgen 
says  that  Bang's  work  on  lymph  glands  gives  support  to  this  con- 
ception, and  throws  doubt  on  the  existence  of  nucleins. 


PHOSPHORUS  METABOLISM 
CLEAVAGE  PRODUCTS  OF  NUCLEOPROTEINS 


17 


Outline  of  the  Processes:  Hydrolytic  cleavages  produce  from 
nucleoproteins,  simple  proteins,  carbohydrates,  phosphoric  acid,  and 
purin  and  pyrimidin  bases,  with  the  intermediate  formation  of  nu- 
cleins  and  nucleic  acids,  as  may  be  represented  by  the  following 
scheme : 

Nucleoproteins 


Proteins 

Proteins 

Carbohydrates 
Pentoses 
Hexoses 
Unidentified 


Nucleins 


Nucleic  acids 


Phosphoric  acid 


Purin  bases      Pyrimidin  bases 
Adenin  Thymin 

Guanin  Cytosin 

Hypoxanthin    Uracil 
Xanthin 


The  carbohydrates  may  appear  as  formic  or  levulinic  acid.  Ox- 
idative processes  transform  hypoxanthin  to  xanthin,  xanthin  to  uric 
acid,  and  sometimes  further  oxidize  the  uric  acid  (probably  allan- 
toin  is  formed  in  such  a  way,  and  perhaps  oxalic  acid  and  urea) . 

PUKIN  BASES 

Four  purins  have  been  obtained,  namely,-  guanin,  adenin,  hypo- 
xanthin and  xanthin.  These  are  spoken  of  also  as  the  nuclein 
bases,  or  alloxur  bases.  The  relations  of  these  to  one  another  and 
to  simple  purin  are  indicated  by  the  following  structural  formulae : 

'Nr^CH  HN— CO  N=C— NH2 

II  II  II 

HC2    5C— HN7  H.N— C  *  C— HN  HC     C-HN 


'CH 


3N—  *C— N* 
Purin 


CH 


N— C— N 
Guanin 


CH 


N— C— N 
Adenin 


HN— CO 

I       I 
HC     C— HN 

\ 


CH 


N— C— N 
Hypoxanthin 


// 


HN— CO 
OC     C— HN 


HN— C— N 

Xanthin 


S 


CH 


18  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Guanin  and  adenin  contain  the  amino  group  (NH2)  while  hy- 
poxanthin  and  xanthin  are  oxygen  derivatives.  The  figures  placed 
beside  members  of  the  purin  ring  are  used  to  designate  the  points 
at  which  substitutions  are  made  in  making  up  the  molecule  of  the 
derivatives,  and  they  may  be  used  in  designating  these  derivatives ; 
thus  guanin  is  2-amino-6-oxypurin,  adenin  is  6-aminopurin,  hypo- 
xanthin  is  6-oxypurin  and  xanthin  is  2,  6-dioxypurin.  By  removal 
of  the  amino  group  guanin  becomes  hypoxanthin,  by  removal  of 
amino  and  introduction  of  oxygen  adenin  becomes  xanthin,  hypoxan- 
thin by  oxidation  becomes  xanthin,  and  by  further  oxidation  this 
becomes  uric  acid,  the  structural  formula  of  which  follows : 

HN— CO 

I  I 

OC     C    — HNX 

II  II  CO 
HN— C    — HN 

Uric  acid  (2,  6,  8 — trioxypurin) 

The  importance  of  these  purin  bases  as  decomposition  products 
of  cell-nuclei  and  of  nuclein  was  shown  by  the  researches  of  Al- 
brecht  Kossel,  and  made  clear  in  his  repeated  discussions.  He  dis- 
covered and  named  adenin  (Kossel,  A.,  1886),  and  we  have  already 
mentioned  the  fact  that  he  marked  the  distinction  between  nucleins 
which  yield  purin  bases  on  cleavage,  and  those  which  do  not,  by 
naming  the  latter  paranucleins.  The  processes  transforming  one 
of  these  purins  to  another  and  the  enzymes  causing  such  trans- 
formations have  received  special  attention  from  Walter  Jones. 

Jones  has  pointed  out  that,  since  guanin  and  adenin  may  so 
readily  be  changed  into  the  other  bases,  it  is  very  probable  that,  at 
least  in  some  of  the  cases,  the  hypoxanthin  or  xanthin  found  among 
the  decomposition  products  was  of  such  origin,  and  was  not  present 
in  the  nucleic  acid  as  such.  By  use  of  his  methods,  which  control 
changes  of  that  kind,  he  was  able  to  prove  that  the  nucleic  acids 
of  thymus,  spleen  and  pancreas  contain  no  other  purin  bases  but 
guanin  and  adenin,  and  he  concluded  that  the  nucleic  acids  from 
these  three  sources,  at  least,  are  identical.  (Jones  and  Austrian, 
1907;  Jones,  1908;  Jones  and  Whipple,  1902).  We  recommend  to 
the  reader  his  clarifying  discussion  (1908)  "On  the  Identity  of 
the  Nucleic  Acids  of  the  Thymus,  Spleen  and  Pancreas,"  in  which 
he  attempts  to  interpret  and  adjust  the  apparently  contradictory 
findings  of  various  authors  as  to  the  identity  of  nucleic  acids  and 
of  their  decomposition  products. 


PHOSPHORUS  METABOLISM  19 

Guanylic  acid  contains  but  one  purin  base,  namely  guanin ;  and 
inosinic  acid,  likewise  only  one,  hypoxanthin. 

PYRIMIDIN  BASES 

Three  pyrimidin  bases  have  been  obtained  from  nucleic  acids,- 
cytosin,  uracil  and  thymin.  The  relations  of  these  to  one  another 
and  to  pyrimidin  are  expressed  in  the  graphic  formulae  as  follows : 

1N=6CH  HN— CNH2  HN— CO  HN— CO 

II  I      II  II  II 

*HC    5CH  OC     CH  OC     CH  OC    C— CH3 

II       II  II  I      II  II     II 

3N— 4CH  N=CH  HN— CH  HN— CH 

Pyrimidin  Cytosin,  Uracil,  Thymin, 

6-amino-2-oxy-         2,  6-dioxy-  5-methyl- 

pyrimidin  pyrimidin  uracil 

The  ring  nucleus  of  these  compounds  is  simpler  than  that  of  the 
purins,  but  might  readily  be  made  from  that  in  case  of  decompo- 
sition, as  may  be  seen  by  inspection  of  the  graphic  formulae. 

Each  of  these  compounds  was  first  prepared  in  Kossel's  labora- 
tory (A.  Kossel  and  A.  Neumann,  1893,  1894 ;  A.  Kossel,  1894 ;  As- 
coli,  1900b),  and  has  been  further  studied  there  as  well  as  else- 
where. 

All  three  of  these  bases  are  in  some  of  the  nucleic  acids,  though 
thymin  and  cytosin  are  most  common.  In  some  cases  where  uracil 
has  been  obtained  it  was  probably  a  secondary  product  derived  from 
cytosin.  By  some  it  has  been  held,  also,  that  all  of  the  pyrimidins 
are  secondary  products  derived  from  purins  (Burian,  1907b) ;  but 
that  idea  seems  to  be  satisfactorily  disproved,  at  least  as  to  its  uni- 
versal application,  by  the  work  of  Steudel  (1905b,  1907e),  and  by 
T.  B.  Osborne  and  Heyl  (1908)  with  regard  to  tritico-nucleic  acid. 
This  last  acid  (from  wheat  embryo)  and  yeast  nucleic  acid  contain 
only  cytosin  and  uracil.  Wheeler  and  Johnson  (1903)  showed  the 
cytosin  from  this  acid  and  that  from  spleen  nucleic  acid  to  be  identi- 
cal. Only  the  simplest  of  the  nucleic  acids,  guanylic  and  inosinic 
acids,  are  without  pyrimidin  bases,  so  far  as  is  known.  The  expres- 
sion thymonucleic  acids  is  sometimes  used  as  a  group  name  for  such 
as  contain  thymin,  and  that  seems  to  be  practically  all  of  those  of 
animal  origin. 

CARBOHYDRATES 

Among  the  cleavage  products  of  all  true  nucleic  acids  or  nu- 
cleins  are  carbohydrates  or  their  derivatives ;  but  the  specific  identi- 
fication of  the  carbohydrate  contained  in  the  nucleic  acid  molecule 


20  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

has  not  generally  been  made.  Kossel  and  many  others  have  found 
both  a  pentose  and  levulinic  acid,  which  latter  is  a  hexose  deriva- 
tive. All  the  thymonucleic  acids  (from  animal  organs)  give  evi- 
dence of  some  kind  of  hexose  (they  yield  levulinic  acid  as  a  hexose 
derivative) ;  while  the  vegetable  nucleic  acids  (yeast  nucleic  acid 
and  triticonucleic  acid  from  wheat  embryos)  and  guanylic  and  in- 
osinic  acids  contain  a  pentose.  Boos  (1909),  however,  in  reinves- 
tigating the  question  of  yeast  nucleic  acid  finds  indication  that  it 
contains  no  pentose.  When  pentose  as  well  as  hexose  has  been  ob- 
tained from  the  acids  derived  from  animal  organs  (Levene  1903a; 
Wohlgemuth,  1903,  1905)  it  may  have  been  due  to  an  admixture  of 
guanylic  acid.  F.  Bauer  (1907)  considers  that  the  pentose  of  inosin- 
ic  acid  is  racemic  arabinose,  while  Neuberg  and  Brahn  (1907)  and 
Haiser  and  Wenzel  (1909)  and  Wohlgemuth  make  it  a  laevo-xylose, 
and  Levene  feels  that  he  disproved  both  of  these  ideas,  and  proved 
that  the  pentose  in  every  case  is  a  dextro-ribose  (Levene  and  Ja- 
cobs, 1909b,  1909c). 

INTERMEDIARY  PRODUCTS 

By  partial  cleavage  several  workers  have  obtained  various  de- 
composition products  from  nucleic  acids  which  are  thought  to  be 
of  especial  interest  as  throwing  light  on  the  makeup  of  the  more 
complex  compound. 

Frequently  the  purin  bases  split  off  first,  but  not  with  sufficient 
ease  to  indicate  that  they  were  in  salt-like  combination.  Alsberg 
(1904)  isolated  what  he  called  heminucleic  acid,  containing  only  one- 
half  of  the  purin  bases,  and  Kossel  and  Neumann  (A.  Kossel,  1894 ; 
A.  Kossel  and  A.  Neumann,  1896)  obtained  an  acid  free  from  all 
the  purin  bases,  without  loss  of  phosphoric  acid,  by  boiling  thymus 
nucleic  acid  with  water.  This  second  acid  was  called  thymic  acid, 
and  it  has  received  considerable  attention,  owing  to  the  suggestion 
that  it  might  bear  the  same  relation  to  paranucleins  that  the  nucleic 
acids  bear  to  true  nucleins.  Milroy  (1896)  proved  that  thymic  acid 
is  not  identical  with  paranucleic  acid  by  showing  that  synthetic 
compounds  of  thymic  acid  with  proteins,  though  much  like  the  nat- 
ural paranucleins,  do  not  give  the  same  acid  on  cleavage.  Steudel 
and  Brigl  (1911)  obtained  the  same,  or  a  similar  thymic  acid,  by 
nitric  acid  oxidation  of  the  thymus  nucleic  acid. 

A.  Kossel  (1894),  A.  Neumann  (1898,  18.99),  and  Kostytschew 
(1903)  found  three  other  modifications  of  nucleic  acid  from  the 
thymus  gland  which  they  called  a-nucleic  acid,  /^-nucleic  acid  and 


PHOSPHORUS  METABOLISM  21 

nucleothyminic  acid,  these  differing  from  one  another  in  the 
amount  of  nuciein  bases  present,  and  showing  certain  differences 
in  properties. 

Alsberg  (1904)  also  obtained,  on  more  energetic  cleavage,  an 
acid  free  from  all  its  purin  bases  and  from  phosphorus,  which  he 
called  nucleotin,  and  which  corresponds  with  Schmiedeberg's  (1900) 
idea  that  the  ground  substance  of  nucleic  acids  is  a  nucleotin-phos- 
phoric  acid  which  is  combined  in  some  way  with  purin  bases.  The 
thymic  acid  of  Kossel  and  Neumann  and  the  thymin-glucophosphor- 
ic  acid  of  Levene  and  Mandel  (1908a)  would  be  such  nucleotin- 
phosphoric  acids. 

Under  the  influence  of  enzyme  action  or  nitric  acid  oxidation 
Steudel  (1908a)-  obtained,  from  nucleic  acid,  products  in  which 
the  sugar  was  in  organic  union  with  phosphoric  acid  after  the  purin 
bases  were  removed.  This  investigation  of  Steudel's  also  made 
evident  that  the  purins  were  closely  bound  to  the  carbohydrate  sec- 
tion of  the  nucleic  acid;  and  the  same  relationship  is  indicated  by 
the  isolation  and  the  synthesis  of  compounds  of  the  carbohydrates 
with  purin  and  pyrimidin  bases  which  have  been  made  by  Levene 
and  his  associates.  Levene,  however,  has  not  succeeded  in  forming 
them  by  such  cleavage  as  that  of  Steudel  mentioned  above. 

The  union  with  the  carbohydrate,  at  least  in  the  case  qf  purin 
bases,  is  said  to  be  glucoside-like,  and  the  compounds  are  called  nu- 
cleosides. Jones  (1911b)  showed  that  enzyme  cleavage  sometimes 
forms  such  bodies  by  splitting  off  phosphoric  acid  from  nucleic 
acids. 

Under  special  conditions  Jones  (1912)  and  Jones  and  Richards 
(1914)  have  obtained  a  simple  nucleic  acid,  guanylic  acid,  from  the 
complex  nucleic  acids  of  pancreas  and  of  yeast. 

Considering  all  of  these  intermediary  products,  it  seems  evi- 
dent that  the  purins  are  more  easily  separated  by  cleavage  than  the 
pyrimidin  bases,  that  the  purin  cleavage  is  gradual,  that  the  phos- 
phoric acid  may  be  as  easily  separated  as  at  least  a  part  of  the  pur- 
in bases,  and  that  the  carbohydrates  never  split  off  so  as  to  leave 
the  purin  and  pyrimidin  bases  united  with  phosphoric  acid.  By 
mild  acid  hydrolysis  phosphoric  acid  is  more  readily  split  off  from 
purin  than  from  pyrimidin  union. 

MONONUCLEIC  ACIDS 

It  is  convenient  to  distinguish  between  nucleic  acids  containing 
only  one  base,  which  may  be  called  mononucleic  acids,  or  simple  nu- 
cleic acids,  and  those  which  contain  more  bases,  and  which  may  be 


22 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


called  polynucleic  acids,  or  complex  nucleic  acids.  The  latter  are 
the  ordinary  nucleic  acids,  and  may  be  looked  on  as  combinations 
of  the  simple  acids  (usually  four  such).  Levene  calls  the  two 
groups,  whether  natural  or  artificial,  mononucleotides  and  poly- 
nucleotides. 

Jones  (1912)  and  Jones  and  Richards  (1914)  have  succeeded 
in  cleaving  a  polynucleotide,  yeast  nucleic  acid,  so  as  to  obtain  a 
mononucleotide,  guanylic  acid,  from  it;  and  the  work  of  Feulgen 
(1913b)  and  of  Knopf  (1914)  suggests  that  Hammarsten's  method 
of  preparing  guanylic  acid  from  the  pancreas  may  involve  a  like 
process. 

INOSINIC   ACID 

An  acid  yielding  on  hydrolysis  only  phosphoric  acid,  hypoxan- 
thin  and  a  pentose,  and  named  inosinic  acid,  is  found  in  meat  ex-_ 
tracts.  It  was  isolated  by  Liebig  (1847),  but  the  formula  then  given 
does  not  include  phosphorus.  It  is  agreed  that  this  is  a  simple  acid 
containing  but  one  molecule  of  each  component.  It  is,  therefore, 
what  Levene  calls  a  mononucleotide.  The  empirical  formula  is 
C10H13N4PO8,  but  several  structural  formulae  have  been  suggest- 
ed. The  work  of  Levene  and  Jacobs  seems  to  have  proved  that  the 
linking  is  such  as  to  include  the  pentoside  inosin,  made  up  of  hy- 
poxanthin  and  the  pentose.  Similar  formulae  are  given  by  Levene 
and  Jacobs  and  by  Haiser  and  Wenzel;  one  which  the  former  au- 
thors suggest  is  given  below,  the  purin  being  bound  to  the  sugar  in 
the  glucoside  manner,  and  the  phosphoric  acid  in  ester  manner: 

H   H    H    H  OC— NH 

HO 
\ 
0=P— O— O—  i  CIL— C— C— C— C  -i N— C    CH 


/ 


HO 


HO    OH 
O  — 


/ 


HC 


^ 


N-C— N 


Phosphoric  acid  Pentose  Hypoxanthin 

(See  Haiser,  1895;  F.  Bauer,  1907;  Neuberg  and  Brahn,  1907; 
Levene  and  Jacobs,  1908,  1909a,  1909b,  1911a;  Haiser  and  Wenzel, 
1909.) 


GUANYLIC    ACID 


Bang  discovered  in  the  pancreas  of  the  ox  a  nucleic  acid  yielding 
no  base  but  guanin,  and  he  named  it  guanylic  acid.  He  at  first 
thought  glycerin  was  present,  but  that  idea  was  not  confirmed.  The 
carbohydrate  found  was  shown  to  be  a  pentose,  and  the  relation  of 


PHOSPHORUS  METABOLISM 


23 


N:P,  that  of  5/.1.  In  his  latest  reports  he  gives  the  formula  C44H65 
N20P4O34,  and  speaks  of  the  molecule  as  made  up  of  4  guanin,  4 
pentose  and  4  phosphoric  acid  groups,  together  with  some  unidenti- 
fied substance  not  containing  nitrogen  or  phosphorus.  (Bang,  1898, 
1901a,  1901b;  Bang  and  Raaschon,  1903;  Bang,  1908,  1910a,  1910b.) 
Steudel  and  Brigl  (1910),  however,  and  Levene  and  Jacobs 
(1909c,  1909e,  1912c)  are  inclined  to  give  to  guanylic  acid  a  formula 
and  structure  more  like  that  of  inosinic  acid.  According  to  them  the 
empirical  formula  is  C10H14N5OsP.  Levene  and  Jacobs  (1912c) 
find  some  reason  to  think  the  structure  is  not  identical  with  that  of 
inosinic  acid,  and  give  the  structural  formula  as  follows: 

OH 


Phosphoric  acid  P — OH 

I      O 

o 

H    H  - 


H— O-CH,— C— C— C— CH 


OH 


o-J 


// 
CH 

\ 


N— C— N 


N— C     C— NH, 


OC— NH 


Pentose  Guanin 

The  phosphoric  acid  is  cleaved  from  guanylic  acid  more  readily 
than  from  inosinic. 

Guanylic  acid  has  been  found  at  least  in  the  pancreas,  the 
spleen,  the  liver  and  milk  glands.  (Steudel  1907d ;  von  Fiirth  and 
Jerusalem,  1907,  1908 ;  Jones  and  Rowntree,  1908 ;  Levene  and  Man- 
del,  1908b;  R.  Odenius,  1899.)  Jones  (1912)  has  produced  it  from 
yeast  nucleic  acid.     See  also  Jones  and  Richards   (1914). 

The  suggestions  of  an  adenylic  acid,  corresponding  to  guanylic, 
have  apparently  been  made  on  insufficient  evidence  (A.  Kossel  and 
A.  Neumann,  1894;  Bang,  1904). 

POLYNUCLEIC  ACIDS 

General  Similarities.  The  numerous  nucleic  acids  of  animal 
origin  containing  both  purin  and  pyrimidin  bases,  which  have  been 
investigated,  show  striking  similarities,  and  very  likely  many  of 
them  are  identical.  It  is  supposed  that  as  a  rule  they  consist  of 
phosphoric  acid,  adenin,  guanin,  2  (or  3)  pyrimidin  bases,  and  a 
hexose,  the  definite  nature  of  which  is  not  determined.    The  plant 


24  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

polynucleic  acids  which  have  been  studied  contain  pentose  in  place 
of  hexose.  Levene  and  LaForge  (Levene,  1909b;  Levene  and 
LaForge,  1910)  find  the  pentose  of  both  yeast  nucleic  acid  and 
triticonucleic  acid  to  be  d-ribose,  and  that  these  acids  yield  the 
same  products  on  partial  hydrolysis.  Therefore  they  think  that 
these  acids  are  identical.  It  is  customary  to  think  of  these  complex 
acids  as  made  up  on  the  basis  of  four  phosphorus  atoms  to  the  mole- 
cule. 

For  convenience,  a  general  list  of  references  to  studies  of  these 
nucleic  acids  is  given  here,  grouped  with  reference  to  their  source. 

NUCLEIC    ACID    STUDIES    ARRANGED    WITH    REFERENCE    TO    THE 
SOURCE    OF  THE  NUCLEIC  ACID 

From  Thymus:  Altmann,  1889;  de  la  Blanchardiere,  1913;  Burian,  1904a, 
1904b,  1907a;  Herlant,  1900;  Iwanoff,  1903;  Jones,  1908;  Jones  and  Austrian, 
1907a;  Kossel,  A.,  1894;  Kossel,  A.,  and  A.  Neumann,  1893,  1894,  1896;  Kosty- 
tschew,  1903;  Levene  and  Mandel,  1908a;  Levene  and  Jacobs,  1912a,  1912b; 
Neumann,  Albert,  1898,1899;  Steudel,  1904,  1905a,  1905b,  1907b,  1907c,  1907d, 
1908a,  1908b,  1912,  1913b;  Tschernorutzky,  Helene,  1912b. 

From  Spermatozoa  or  Sperm:  Alsberg,  1904;  Altmann,  1889;  Herlant, 
1900;  Inouye,  1904,  1906;  Kossel,  A.,  1896;  Levene  and  Mandel,  1906e;  Miescher, 
1878,  1896,  1897;  Noll,  1898;  Sauerland,  1910;  Schmiedeberg,  1900;  Steudel, 
1906a,  1906b,  1907b,  1907c,  1907d,  1911a. 

From  Spleen:  Bang,  1903,  1904;  Inouye,  1904;  Jones  1908;  Levene,  1901b, 
1903a,  1903b,  1904,  1905;  Levene  and  Mandel,  1906a. 

From  Pancreas:  Feulgen,  1913b;  Jones,  1908;  Levene,  1901b,  1903a,  1903b, 
1903d,  1904;  Levene  and  Jacobs,  1909c,  1909e;  Sauerland,  1910;  Steudel,  1907f; 
von  Fiirth  and  Jerusalem,  1907. 

From  Testes:     Inouye,  1904;  Levene,  1903f,  1904. 

From  Fish  Roe:  Levene,  1901b;  Levene  and  Mandel,  1906b;  J.  A.  Mandel 
and  Levene,  1906b;  Tschernorutzky,  Helene,  1912a. 

From  Yeast:  Altmann,  1889;  Ascoli,  1899,  1900b;  de  la  Blanchardiere, 
1913;  Boos,  1906,  1909;  Burian,  1904a,  1904b,  1907a;  Herlant,  1900;  Jones,  1912; 
Jones  and  Richards,  1914;  Kowalevsky,  1910';  Levene,  1901b,  1903d,  1909b;  Le- 
vene and  Jacobs,  1909c,  1909f,  1909g,  1910,  1911b;  Levene  and  La  Forge,  1912; 
Liebermann,  1888b,  1889,  1890;  Liebermann  and  Bitto,  1893;  Tschernorutzky, 
Helene,  1912b. 

From  Wheat  Embryo:  Levene  and  La  Forge,  1910;  Osborne,  T.  B.,  and 
Campbell,  1900a;  Osborne,  T.  B.,  and  Harris,  1902;  Osborne,  T.  B.,  and  Heyl, 
1908;  Wheeler  and  Johnson,  1903. 

From  Brain:     Levene,  1899,  1903c,  1903f,  1904. 

From  Liver:     Levene,  1903e;  Levene  and  Jacobs,  1909c. 

From  Kidney:  J.  A.  Mandel  and  Levene,  1906a. 

From  Intestinal  Tissue:     Araki,  1903b;  Inouye  and  Kotake,  1905. 

From  Mammary  Glands:  Basch,  1898;  Lobisch,  1906;  J.  A.  Mandel  and 
Levene,  1905. 

From  Human  Placenta:    Kikkoji,  1907b. 

From  Leucocytes:     Ascoli,  1900a;  A.  Kossel,  1893. 

General  Work:  Levene,  1903c,  1910;  Levene  and  Medigreceanu,  1911b, 
1911c,  1911d;  Schmiedeberg,  1907. 

Elementary  Composition.  Since  these  compounds  are  in  them- 
selves so  complex,  and,  moreover,  always  exist,  whether  in  the  body 
or  in  the  food,  in  conjunction  with  undetermined  proportions  of 


PHOSPHORUS  METABOLISM 


25 


other  compounds,  and  since  it  is  quite  doubtful  whether  they  have 
ever  been  examined  either  in  a  pure  condition  or  in  their  natural 
state,  it  seems  that  the  percentage  content  of  phosphorus  or  other 
constituents  has  no  especial  significance  other  than  as  contributing 
to  a  knowledge  of  the  formulae,  and  then  only  as  an  aid  to  the  study 
of  the  structure  as  indicated  by  chemical  reactions  and  cleavage 
products.  The  question  of  the  percentage  composition  of  the  proteins 
which  are. united  to  the  nucleic  acids  is  practically  untouched.  On 
account  of  such  interest  as  they  may  possess,  a  few  empirical  form- 
ulae of  nucleic  acids  are  presented.  They  are  determined  in  part 
by  analysis    and  in  part  by  probable  structure. 

FORMULAE  OF  A  FEW  NUCLEIC  ACIDS 


Nucleic  acid 

Investigator 

Date 

and 

reference 

Empirical 
formula 

Schmiedebergf 
Steudel 
Levene 
Kowalevsky 
Osborne  and  Harris 

1900 

1912 

1909b 

1910 

1902 

C40H56N14P4O26 

C43H61N15P4034 

C38H49N15P4029 

C29H42N13P3023 

Triticonucleic  acid,  (from  wheat  embryo)    

C41H61N16P403I 

Structure.  Two  extensive  investigations  which  have  led  to 
somewhat  definite  pictures  of  the  structure  of  these  nucleic  acids 
are  those  of  Steudel  and  of  Levene  and  his  collaborators.  With 
regard  to  the  principal  thymus  nucleic  acid  (and  Steudel  thinks  that 
from  herring  sperm  is  identical  with  this)  these  authorities  are 
agreed  that  the  molecule  contains  the  four  bases  guanin,  adenin, 
thymin  and  cytosin  in  unimolecular  relation,  and  each  bound  to  a 
hexose  molecule  which  in  turn  is  bound  to  phosphoric  acid,  there 
being  four  phosphoric  acid  molecules ;  also  that  the  linking  of  the 
hexose  with  the  purin  bases  (and  perhaps  that  with  pyrimidin 
bases)  is  glucoside-like,  but  the  union  with  pyrimidins  is  much  less 
easily  broken  than  that  with  purins.  Steudel's  picture  involves  a 
more  condensed  form  of  phosphoric  acid  than  Levene's.  Levene, 
applying  the  name  mononucleotide  to  the  individual  complexes  of 
a  single  base,  carbohydrate  and  phosphoric  acid,  which  he  considers 
quite  analogous  to  inosinic  and  guanylic  acids  (see  the  graphic 
formulae  given) ,  looks  upon  these  acids  as  polynucleotides  made  up 
of  four  mononucleotides.  Jones  (1912)  has  brought  about  a  cleav- 
age of  yeast  nucleic  acid  with  the  production  of  the  mononucleotide, 
guanylic  acid.  Steudel's  formula  does  not  attempt  to  represent  the 
linking  within  the  hexose,  nor  in  the  base-hexoside. 


26  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Steudel's  (1912)  formula  for  thymus  acid  is: 

(OH)2=  P— C„HuN5Od  (Guanin-hexoside) 

/\ 

O  O 

\/ 

P — C10H14N3O6  (Cytosin-hexoside) 

/\ 

O  O 
\/ 

P— CnH15N207  (Thymin-hexoside) 

/\ 
O  O 
\/ 
(OH)2=  P— CuH14N505  (Adenin-hexoside) 

In  Levene  and  Jacobs's  formula  (p.  27)  the  two  pyrimidin-nu- 
cleotides  are  represented  as  linked  to  each  other  by  their  sugar, 
while  the  phosphoric  acids  of  the  purin-nucleotides  serve  to  link  the 
purin-nucleotides  with  the  sugar  (not  the  phosphoric  acid)  of  the 
pyrimidin-nucleotides.  These  conceptions  correspond  with  the  in- 
termediary products  obtained.* 

From  yeast  nucleic  acid  Levene  and  Jacobs  (1909f,  1909g,  1910, 
1911b)  find  the  phosphoric  acid  more  readily  cleaved  and  the  purins 
less  readily  than  from  thymus  nucleic  acid,  and  the  structural  form- 
ula suggested  is : 

OH 

/ 
0=P-C5H804C5H4N5 

\ 
O 

/ 
0=P-C5H804C5H4N50 

O 

/ 
0=P— C5H804-  C4H4N30 

O 
/ 
0=P-C5H804C4H3N208 
\ 

OH 


PHOSPHORUS  METABOLISM 


27 


Levene  and  Jacob's  (1912b)  formula  for  thymus  nucleic  acid  is: 

O  — 


H.    H 

I        I 
H2C  —  C  —  C  —  C  —  C  —  C  —  Guanin 

I      !      I      I      I      I 

■  i    OH     H  OH  OH     H 
O 
I 
HO— P=0  OH 

,  / 

p=o 

o 


o 


OH 


O 


H 

I 


H 


H2C-C— C  —  C  — ( 

1      1        i 

:— c- 

i 

-  Thymin 

1      1        1 
H    H     OH 

( 

i 

H 

H    H     OH 

1       1        1 

H 

j 

1       1        1 
H2C  — C— C— C  — ( 

:— c- 

-  Cytosin 

o 


H     H 

— o— 


o 


OH 


/ 

PrO 

\ 
HO— P=0      OH 

O 

|  OH    HOH  OH     H 

H9C  — C  — C—  C  —  C  —  C  —  Adenin 


H 


H-   H 
O 


Levene's  ideas  as  to  the  structure  and  cleavage  of  nucleic 
acids,  together  with  the  distribution  of  cleaving  enzymes  in  the 
animal  body  were  clearly  summarized  by  Levene  and  Medigreceanu 
(1911d) .      See  Ferments  of  Individual  Organs. 

Helene  Tschernorutzky  is  making  a  study  of  the  glucoside  re- 
lation in  these  acids  by  attempting  cleavage  by  means  of  enzymes 
which  are  known  to  have  the  power  to  split  glucosides.    So  far  the 


28  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

evidence  is  unfavorable,  for  the  work  reported  in  August,  1912 
(Tschernorutzky,  H.,  1912b)  resulted  in  a  separation  of  inorganic 
phosphate  as  well  as  purin  bases,  indicating  a  more  thoroughgoing 
cleavage,  a  nuclease  action. 

Burian  (1904a,  1904b,  1907a)  believes  the  purin  bases  to  be 
linked  to  the  remainder  of  the  nucleic  acid  at  the  7-place  of  the 
purin  ring.  Levene  thinks  that  the  pyrimidins  are  linked  at  either 
the  8-  or  the  4-  place.    (Levene  and  LaForge,  1912). 

ENZYMES  CONCERNED  IN  NUCLEIC  ACID  CLEAVAGE 

The  name  nuclease  has  been  used  to  signify  an  enzyme  which 
brings  about  complete  disruption  of  the  nuclein  molecule.  It  was 
first  applied  by  Iwanoff  (1903).  Evidence  of  such  an  enzyme  (or 
enzymes)  has  repeatedly  been  found  in  aqueous  extracts  of  animal 
organs  (Jones,  1904a,  1904b;  Levene  and  Medigreceanu,  1911b, 
1911d) ;  but  it  is  probable  that  the  process  is  due  to  several  enzymes 
acting  in  succession. 

Jones  distinguishes  four  enzymes  having  the  power  to  deamin- 
ize,  two  of  which,  guanase  (Jones  and  Partridge,  1904)  and  adenase 
(Jones  and  Winternitz,  1905),  act  directly  on  the  purin  bases 
(guanin  and  adenin,  respectively),  and  two  on  the  glucosides  of 
these  bases  (guanosin-deaminase  converting  guanosin  to  xanthosin, 
and  adenosin-deaminase  converting  adenosin  to.  inosin,  the  hypo- 
xanthin  glucoside)  (Jones,  1911b).  Accordingly  he  recognizes  two 
general  paths  by  which  the  nucleic  acid  may  be  transformed  for 
the  formation  of  uric  acid;  (1)  a  nuclease  may  cleave  off  the  phos- 
phoric acid  and  the  carbohydrate,  leaving  the  purin  (and  the  pyri- 
midin)  bases,  the  purin  bases  then  being  transformed  by  the  de- 
aminizing  ferments  guanase  and  adenase  into  xanthin  and  hypoxan- 
thin,  respectively,  the  hypoxanthin  being  oxidizable  to  xanthin,  and 
xanthin  to  uric  acid  through  the  agency  of  xanthooxidase;  or  (2) 
the  nucleic  acid  may  first  be  broken  down  by  a  cleavage  of  phos- 
phoric acid  only,  by  which  guanosin  and  adenosin  are  left,  and  they 
may  be  deaminized  at  once  into  xanthosin  and  inosin,  and  then  have 
their  carbohydrates  cleaved  off,  with  the  formation  of  xanthin  and 
hypoxanthin ;  or  the  carbohydrates  may  break  off  before  deaminiza- 
tion,  which  would  transform  them  to  guanin  and  adenin,  upon  which 
guanase  and  adenase  act  as  in  the  other  line  of  cleavage ;  in  either 
case  the  xanthooxidase  completes  the  process.  The  following  dia- 
gramatic  scheme  taken  from  Amberg  and  Jones  (1911b)  may  help 
to  make  this  clear.  The  arrows  indicate  possible  changes,  for  each 
of  which  a  distinct  ferment  might  be  required.  The  numbered  ones 
have  been  proved  to  be  present  in  one  or  more  organs  or  fluids. 


PHOSPHORUS  METABOLISM 


29 


DIAGRAMATIC  SCHEME  OF  POSSIBLE  CLEAVAGES  OF  NUCLEIC  ACID 

AND  THE  TRANSFORMATIONS  RESULTING  IN  THE 

FORMATION  OF  URIC  ACID 

Nucleic  acid,  shown  as  dinucleoside 

OH  OH 

I  / 

0=P— 0-C5H803-C5HN4 

\  \ 

O  NH2 

/ 
O  =  P-O  •  C5Hg03  •  C5H2N4(NH2) 

I 
OH 


OH  OH 

/  / 

C5H2N4  C5HN4  •  C5H904    CJH,Nt(NH,)-C!H,01       C5H3N4(NH2) 

\  \ 

NH,  NH2 


Xanthosin 
OH 
/ 


Inosin  __ 


C5HN4  •  C5H904        C5H2N4(OH)-C5H904 


\ 


OH 


Xanthin    <- 
C5H2N4— OH 
\ 
OH 


->       Hypoxanthin 
C8H3N4(OH) 


Uric  acid 

OH 

/ 

C5HN—  OH 

\ 
OH 


30  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Enzymes  represented: 

1.  Phosphonuclease — cleaving  off  phosphoric  acid. 

2.  Purin  nuclease — cleaving  off  purins. 

3.  Guanosin  deamidase — removing  amino  group  (NH2)  of 
guanosin. 

4.  Adenosin  deamidase — removing  amino  group  (NH2)  of 
adenosin. 

5.  Adenase — removing  amino  group  of  adenin. 

6.  Guanase — removing  amino  group  of  guanin. 

7.  Xanthosin  hydrolase — hydrolyzing  xanthosin,  removing 
carbohydrate. 

8.  Inosin  hydrolase — hydrolyzing  inosin,  removing  carbohy- 
drate. 

9.  Xanthooxidase — oxidizing  hypoxanthin  or  xanthin  to  uric 
acid. 

By  the  terminology  of  Levene  and  Medigreceanu  (1911d)  such 
nucleic  acids  as  the  above  would  be  called  "dinucleotides,"  the 
simplest  of  polynucleotides,  and  the  first  enzymatic  process 
(brought  about  by  "nucleinases")  is  the  cleavage  of  polynucleotides 
to  simple  nucleotides.  The  enzymes  cleaving  these  nucleotides,  such 
as  Jones  calls  phosphonucleinase,  Levene  and  Medigreceanu  call 
nucleotidases,  and  the  glucosides  resulting  they  call  "nucleosides." 
The  enzymes  causing  the  cleavage  of  nucleosides  are  named  "nucleo- 
sidases." 

STUDIES    ON    ARTIFICIAL    NUCLEIN    SYNTHESIS 

Nucleins  have  not  been  made  artificially.  A  number  of  workers 
have,  however,  succeeded  in  bringing  about  synthesis  of  organic 
compounds  of  phosphoric  acid  which  seem  to  resemble  in  their 
make-up  fractions  of  the  nuclein  molecule,  and  certain  of  them  have 
been  called  nucleins.  None  of  these,  so  far  as  we  have  learned,  are 
unions  of  the  phosphoric  acid  with  both  carbohydrate  and  nitrogen- 
ous components,  and  therefore  they  have  not  even  all  the  constitu- 
ents of  the  nucleic  acids.  The  processes  which  have  been  observed 
may  be  looked  on  as  partial  syntheses  of  nucleins,  for  phosphoric 
acid  unions  have  been  made  with  carbohydrates  on  the  one  hand, 
and  with  simple  proteins  on  the  other. 

Apparently  direct  union  of  simple  proteins  with  phosphoric 
acid  radicals  occurred  in  the  studies  of  Pohl  (1889),  of  Liebermann 
(1888b),  of  Fuld  (1902a),  and  perhaps  of  Malfatti  (1892,  1893). 
In  all  of  these  cases  metaphosphoric  acid  or  its  salt  was  used.  Bech- 
hold  (1901)  reports  a  similar  compound  of  orthophosphoric  acid 
with  albumin  formed  by  use    of    phosphorus    oxychloride.      Such 


PHOSPHORUS  METABOLISM  31 

compounds  as  these  were,  at  the  time  they  were  made,  looked  upon 
as  synthetic  nucleins  (or  paranucleins) .  Giertz  (1899)  definitely 
proved  that  in  such  a  compound  as  Liebermann's  the  phosphoric  acid 
does  not  bear  the  same  relation  to  the  albumin  that  it  does  in  the 
natural  paranuclein  from  casein.  We  now  know  that  even  the  para- 
nucleins are  not  made  up  simply  of  phosphoric  acid  and  albumin. 

Nucleic  acids  readily  precipitate  proteins  from  solution,  and, 
according  to  the  investigation  of  Milroy  (1896),  there  are  then 
formed  firm  chemical  compounds  which  somewhat  resemble  nu- 
cleins. Those  formed  from  thymic  acid  and  protein  were  not  identi- 
cal with  natural  paranuclein. 

By  methods  similar  to  that  of  Bechhold,  but  carried  out  at  low 
temperatures,  Neuberg  and  his  associates  (Neuberg  and  Pollak, 
1910a,  1910b,  1910c,  1910d ;  Neuberg  and  Kretschmer,  1911)  have 
succeeded  in  forming  phosphate  compounds  with  the  carbohydrates 
saccharose,  glucose,  fructose,  maltose,  lactose  and  galactose,  with 
glycerin  and  also  with  the  phosphorus-free  proteins  lactalbumin  and 
serum  globulin.  These  phosphorized  proteins  are  said  to  resemble 
casein  in  elementary  composition  and  in  behavior,  in  so  far,  at  least, 
that  the  first  cleavage  produced  in  tryptic  and  in  peptic  digestion 
seems  to  correspond  to  the  digestion  of  casein  as  described  by  Sal- 
kowski  (1899,  1901).  The  authors  consider  the  phosphoric  acid  to 
have  entered  into  the  protein  molecule  by  uniting  with  the  amino- 
or  imino-  group  of  the  protein  to  form  a  substituted  phosphaminic 
acid. 

Carbohydrate-phosphoric  acid  esters  have  frequently  been  pro- 
duced in  recent  years  in  connection  with  yeast  fermentation  of  sug- 
ars, the  union  apparently  being  effected  by  a  ferment  present  in  the 
yeast.  These  complexes  act  as  catalytic  agents  in  accelerating  the 
fermentation  by  yeast  or  yeast-juice,  and  it  is  thought  that  their 
formation  and  subsequent  hydrolysis  furnish  the  mechanism  by 
which  the  well-known  favorable  influence  of  phosphates  in  such  fer- 
mentation is  exerted.  Von  Lebedew  and  Griaznoff  (1912)  give  a 
definite  picture  of  the  process  as  they  conceive  of  it.  The  extensive 
work  of  Young  and  Harden,  of  von  Lebedew,  and  of  Euler  and  his 
associates  is  especially  to  be  noted.  (See  Wroblewski,  1901 ;  Iwanoff, 
1907;  A.  Harden  and  Young,  Proc.  Chem.  Soc.  21  (1905),  189;  A. 
Harden  and  Young,  1906,  1908a,  1908b,  1909,  1910,  1911a,  1911b; 
Young,  1907,  1909,  1911 ;  A.  Harden,  1910 ;  A.  Harden  and  Norris, 
1910;  von  Lebedew,  1910a,  1910b,  1911a,  1911b,  1911c;  Euler  and 
Fodor,  1911;  Euler  and  Lundeqvist,  1911;  Euler  and  Kullberg, 
1911a,  1911b;  Euler  and  Ohlsen,  1911,  1912;  Harden  and  Young, 
1912 ;    Euler,    1912a,    1912b,    1913 ;    Euler    and  Backstrom,  1912 ; 


32  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Euler  and  Funke,  1912;  Euler  and  Johansson,  1912a,  1912b, 
von  Lebedew,  and  Griaznoff,  1912.)  Young  identified  his 
compound  as  a  diphosphoric  acid,  ester  of  the  hexose  fruc- 
tose, and  others  confirm  this  finding  of  a  diphosphoric 
ester.  Euler  has  given  much  attention  to  the  enzyme  involved, 
which  he  names  "phosphatese."  Euler  and  Johansson  (1912b)  find 
that  neither  glycerol,  inositol,  mannitol  nor  alanine  forms  such  an 
ester  with  phosphates,  nor  do  glucose,  fructose  and  mannose  till 
after  they  have  been  altered  in  some  way  by  an  enzyme  or  by  dilute 
alkali.  Von  Lebedew  and  Griaznoff  find  that  dioxyacetone  forms  an 
ester  with  phosphate  but  glycerin  aldehyde  does  not.  According  to 
their  conception  the  yeast  fermentation  of  glucose  first  cleaves  the 
sugar  into  these  two  compounds  and  then  the  latter  ferments  direct- 
ly; the  former  by  way  of  the  intermediate  phosphate  ester  forma- 
tion. 

Langheld  (1910,  1911,  1912)  has  synthesized  first  meta-  and 
from  them  orthophosphoric  acid  esters  from  the  alcohols.  Among 
his  products  is  a  monoester  of  fructose  which  he  thinks  may  be 
identical  with  the  hexose  compound  of  Young  and  of  von  Lebedew. 

These  synthetic  investigations  will  probably  give  valuable  evi- 
dence, though  indirect,  as  to  the  structure  of  the  natural  organic 
phosphorus  compounds  and  as  to  the  anabolic  processes  which  may 
go  on  in  the  animal  body. 

THE  PHOSPHORUS  OF  NUCLEOPROTEINS 

Probably  all  of  the  phosphorus  of  the  nucleoproteins  is  con- 
tained in  the  nucleic  acid  fraction,  and  as  has  been  shown,  it  is  there 
present  in  a  highly  oxidized  state  and  apparently  ready  formed  as 
phosphoric  acid,  more  or  less  dehydrated,  or  else  as  esters  of  ortho- 
phosphoric  acid.  If,  therefore,  the  natural  substances,  the  nucleo- 
proteins, are  completely  digested,  they  yield  phosphoric  acid;  and 
if  only  partially  digested,  the  phosphorus  is  left  in  the  form  of  sub- 
stances, nucleins  or  nucleic  acids,  which  may  or  may  not  be  of  direct 
use  in  the  body.  The  question  of  the  extent  to  which  digestive 
cleavage  does  take  place  will  be  considered  in  the  discussion  of  met- 
abolism of  nucleoproteins. 

CHEMISTRY  OF  PHOSPHOPROTEINS 

CASEIN 

Our  treatment  of  the  literature  on  the  chemistry  of  casein  is 
by  no  means  exhaustive.  Our  earliest  note  is  of  a  discussion  by 
Braconnot  in  1830.  Since  that  date  there  have  been  probably  a 
thousand  articles  on  this  subject.  For  general  reviews  see  Fuld 
(1902b)  andRaudnitz  (1903). 


PHOSPHORUS  METABOLISM  33 

NATURE  OF  THE  COMPOUND 

Casein  is  a  phosphoprotein.  The  recommendations  made  in 
1908  by  the  joint  committee  of  the  American  Physiological  Society 
and  the  Society  of  Biological  Chemists  place  phosphoproteins  among 
the  conjugated  proteins  and  define  them  as  "Compounds  of  the  pro- 
tein molecule  with  some,  as  yet  unidentified,  phosphorus-containing 
substance  other  than  a  nucleic  acid  or  lecithins,"  adding  "they  are 
possibly  esters  of  some  phosphoric  acid  or  acids  and  protein."  The 
earlier  view  which  looked  on  them  as  simple  proteins  and  used  the 
names  nucleoalbumin  or  phosphoglobulin  is  thus  discarded. 

Plimmer  (1913b),  judging  from  the  comparative  instability 
toward  alkali  hydrolysis,  thinks  that  the  phosphoproteins  are  not 
esters,  and  that  the  phosphorus  is  probably  combined  with  one  of 
the  amino-acids. 

Phosphoproteins  differ  from  nucleoproteins  in  not  yielding 
purin  bases  on  cleavage,  and  therefore  not  containing  true  nucleic 
acids.  Incomplete  peptic  digestion  does,  however,  leave  a  phos- 
phorized  body  resembling  the  nucleins,  which  are  derived  in  the 
same  way  from  nucleoproteins.  The  residues  from  phosphoproteins 
are  called  para-  or  pseudonucleins.  All  the  attempts  that  have  been 
made  to  isolate  a  paranucleic  acid  from  paranuclein  have  failed  in 
that  the  product  obtained  still  contained  protein.  Levene  and  Als- 
berg  (1901)  reported  such  an  acid  from  the  paranuclein  of  ovovitel- 
lin, as  also  did  Salkowski  (1901),  Reh  (1908)  and  M.  Dietrich 
(1909)  from  the  paranuclein  of  casein.  Salkowski  examined  his 
preparation  in  the  form  of  an  iron  compound,  Reh  as  a  uranyl  com- 
pound, and  Dietrich  separated  four  calcium  preparations.  Reh 
thought  his  compound  corresponded  with  a  uranyl  ammonium  phos- 
phate in  which  the  ammonium  was  replaced  by  an  albumose  com- 
plex bound  in  a  manner  more  or  less  ester-like  but  not  like  a  salt,  and 
he  calls  it  a  polypeptid  phosphoric  acid.  The  formula  given  for  the 
uranyl  ester  is  C29H5CNsP2U20,4.  Dietrich  speaks  of  his  com- 
pound as  a  phosphorus-containing  casein  peptone. 

OCCURRENCE  OF  CASEIN 

Casein  occurs  only  in  the  milk  of  mammals.  It  forms  the  main 
part  of  the  curd  of  milk.  (For  table  of  casein  contents  of  the  milk 
of  different  species  consult  index.)  The  amount  in  cow's  milk 
is  usually  given  as  not  far  from  3  percent,  while  that  of  human  milk 
is,  by  most  observers,  put  as  low  as  1  percent,  or  even  lower.  In 
this  respect  the  milks  of  the  goat,  pig  and  elephant  seem  to  be 
nearly  like  that  of  the  cow,  those  of  the  ass  and  mare  nearly  like 
that  of  woman,  while  those  of  the  buffalo,  dog,  cat,  ewe,  and  guinea 
pig  have  higher  casein  content  than  that  of  the  cow. 


34 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


In  milk  the  casein  is  in  suspension  in  combination  with  calcium 
and  perhaps  calcium  phosphate.  According  to  Lindet  (1913a, 
1913b)  there  are  two  caseins  in  milk  which  may  be  distinguished 
by  their  action  on  polarized  light.  The  solubility  of  both  in  the 
milk  is  favored  by  the  presence  of  phosphates  and  citrates. 

ELEMENTAEY    ANALYSIS    AND    EMPIRICAL    FORMULA 

The  table  below  shows  the  elementary  analysis  of  casein  from 
cow's  milk  as  reported  by  different .  authors  and  the  second  table 
similar  analyses  of  casein  from  the  milk  of  different  species. 

ELEMENTARY  ANALYSIS  OF  CASEIN  FROM  COW'S  MILK— Percent 


Author 
quoted 

Date 
and 
refer- 
ence 

C 

H 

N 

S 

P 

0 

Makris 

1876 

53.622 

7.422 

14.20 

Hammar- 
sten 

1883, 
1885a 

52.96 

7.05 

15.65 

0.758 

0.847 

22.78 

Mean      results 
from  many 
substances. 

Chitten- 
den   and 
Painter 

1885 

0.88 
0.85 

Seven  samples 

Beohamp 

1893 

0.752 

Mean  of  four 
determinations 

Von 

Szontagh 

1893 

0.87 

Hemp  el 
(Leh- 

m  arm's 

1894 

54.00 

7.04 

15.6 

0.771 

0.847 

Computed  to 
ash-free    basis 

work  ) 

Wr6blew- 
ski 

1894a, 
1894b 

53.00 

7.00 

15.70 

0.80 

0.85 

25.65 

Osborne, 
Thomas 

1902 

0.86 

Storch 

1902 

53.00 

6.4 

15.7 

0.80 

0.85 

22.65 

Quoted    by 

Ellen- 
berger 

1902 

53.07 

7.13 

15.64 

0.76 

0.8 

Mann,  Chem- 
istry of  Pro- 
teids 

Burow 

1905 

52.825 

7.095 

15.64 

0.725 

0.808 

22.906 

Taken  through 
Tangl,    1908 

Tang] 

1908 

52.69 

6.81 

15.65 

0.832 

0.877 

23.14 

Kikkoji 

1909 

0.86  + 

Van 

Slyke  and 
Bosworth 

1912, 
1913a 

53.50 

7.13 

15.80 

0.72 

0.71 

22.08 

Ash  only  0.06 
percent 

PHOSPHORUS  METABOLISM 


35 


ELEMENTARY.  ANALYSIS  OF  CASEIN  FROM  DIFFERENT  KINDS 

OF  MILK— Percent 


Source  of 
milk 


Author  quoted 


reference 

C 

H 

N 

S 

P 

53.00 

7.00 

15.7 

0.'8 

0.86 

1876 

52.353 

7.266 

14.650 

(  1894a, 
1  1894b 

52.24 

7.325 

14.97 

1.117 

0.679 

j  1894a, 
( 1894b 

52.41 

7.28 

15.03 

1.11 

0.651 

1908 

52.82 

7.04 

14.47 

0.78 

0.26 

1910 

52.51 

7.07 

14.61 

0.731 

0.243 

1908 

52.88 

7.81 

15.78 

0.833 

0.773 

1902 

54. 

7.0 

14.4 

0.84 

1.04 

1908 

52.57 

7.01 

16.28 

0.588 

1.057 

1908 

52.92 

7.05 

15.71 

0.717 

0.809 

1905 

52.805 

7.02 

15.64 

0.718 

0.815 

1908 

52.90 

6.86 

15.48 

0.700 

0.760 

1905 

52.866 

7.045 

15.595 

0.733 

0.810 

1908 

52.36 

7.09 

16.44 

0.528 

0.871 

Cow 
Woman 


Buffalo 

Ass 

Ass 

Sheep 

Goat 

Goat 

Rabbit 

Mare 


Summarized  from  previous 

table 

Makris 

Wroblewski1 

Wrohlewski2 

Bergrell  and  Lang-stein3. . . 
Langstein  and  Edelstein4. 

Tangl 

Storch 

Tangl 

Tang-K 

Burow5 , 

Tang-1 

Burow5 

Tangl 


22.8 
23.66 


21.925 

23.32 

22.495 

22.794 

23.002 

23.300 

22.952 

22.705 


(1)  Mean   of   eight   analyses   on   five   preparations. 

(2)  Mean  of  six  analyses  on  three  whitest  preparations. 

(3)  Mean  of  two.       Hot  water  washing  may  have  removed  some  S  and  P. 

(4)  Mean  values  for  five  preparations. 

(5)  Taken  from  Tangl,  1908. 

From  Hammarsten's  analytical  data  Richmond  (1901)  com- 
puted the  "probable  approximate  formula"  for  casein  as  separated 
from  cow's  milk  by  Hammarsten's  method,  C162H258N41SP052,  and  for 
the  salt  separable  by  the  porcelain  mter,C162H255N41SP052CaNa.I/2 
(Ca3P208).  The  formula  given  by  T.  Osborne  (1902)  is  C708H1130 
N180S4P4O224,  indicating  a  more  condensed  molecule.  Raudnitz 
(1903)  quotes  formulae  of  the  simple  protein  from  Knop,  C64H100 
N1602  and  from  Millon-  Commaile,  C10SH97N14O29. 


PHYSICAL    AND    CHEMICAL    PROPERTIES    OF    CASEIN 

Casein  is  a  white,  amorphous  powder,  practically  insoluble  in 
water.  It  is  an  acid  and  as  such  readily  dissolves  in  solutions  of 
the  hydroxides  or  the  carbonates  of  alkalis  or  alkaline  earths  by 
forming  soluble  salts.  These  salt  formations  have  been  much  in- 
vestigated. (See  Soldner,  1888;  Bechamp,  1893;  Courant,  1891; 
de  Jager,  1897 ;  Osborne,  W.  A.,  1901-02 ;  Long,  1906a ;  Robertson, 
1906-07,  1908;  VanSlyke  and  Bosworth,  1912,  1913b,  1913c,  1913d; 
Bosworth  and  VanSlyke,  1913).  L.  L.  VanSlyke  and  A.  W.  Bos- 
worth (VanSlyke  and  Bosworth,  1912,  1913c;  Bosworth,  1913)  de- 
cide that  it  has  8  valences  toward  metals,  and  that  its  molecular 
weight  is  8888.  The  specific  rotation  of  alkali  solutions  of  casein 
has  been  investigated  by  Long  (1905,1906a). 


.  36  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Being  a  protein,  casein  also  forms  union  with  acids,  and  ion- 
protein  compounds  with  salt  ions.  Long  (1907b)  discusses  the 
combining  power  toward  acids.  The  alkali  or  alkaline  earth  solu- 
tions of  casein  are  precipitated  by  a  very  little  acid,  and  are  soluble 
in  excess  of  the  acid  (in  most  cases).  Increase  of  temperature 
favors  the  precipitation  by  acids.  The  presence  of  certain  salts 
is  said  to  favor  the  solubility  of  casein  and  its  salts,  but  sodium 
chloride,  magnesium  sulphate  and  some  others  "salt  out"  casein 
from  solution.  (See  Biedert,  1887;  Schroter,  1887;  Storch,1897, 
1902 ;  Schmidt-Nielsen,  1906;  Robertson,  1906-07.) 

According  to  Dakin  and  Dudley  (1913a,  1913b),  dilute  alkalis 
acting  on  casein  at  low  temperatures  racemize  the  casein  by  tauto- 
meric changes  within  the  protein  complex.  Such  racemized  casein, 
and  racemized  caseose  resulting  from  it  by  hydrolysis,  were  found 
to  escape  digestion  and  absorption  when  given  either  per  os  or  sub- 
cutaneously  to  a  dog. 

The  most  characteristic  property  of  casein  is  its  coagulation 
with  rennet  in  the  presence  of  lime  salts.  Hydrolysis  by  boiling 
acids,  or  under  the  influence  of  enzymes,  cleaves  casein  as  it  does 
other  proteins.  Buxton  and  Rake  (1911)  discuss  the  various 
types  of  coagulation  in  connection  with  their  study  of  the  flocking 
out  of  colloids  in  general. 

COAGULATION   BY   ACIDS   AND   BY   HEAT 

Pure  casein  salt  solutions  and  fresh  milk  do  not  coagulate  on 
boiling,  but  in  the  presence  of  free  acid  coagulation  may  take  place 
below  the  boiling  temperature.  The  coagulum  formed  in  the  case 
of  milk  includes  fat  and  calcium  phosphate.  The  slight  pellicle 
which  coats  over  milk  when  it  is  warmed  is  of  the  same  composition. 

The  usually  accepted  explanation  of  the  precipitation  of  casein 
by  acids  is  that  the  casein  is  held  in  solution  by  chemical  union  with 
a  base  (lime  in  the  case  of  milk) ,  that  added  acid  removes  the  base, 
allowing  the  insoluble  casein  to  precipitate,  and  that  excess  of  acid 
unites  with  casein  forming  a  compound  which  is  more  or  less  readily 
soluble.  Probably  what  actually  occurs  is  not  quite  so  simple; 
it  may  be  rather  that  there  are  salts  of  different  proportions  of  base 
formed  in  the  presence  of  different  proportions  of  acid,  and  that 
some  of  these  salts  are  soluble  and  some  are  insoluble.  L.  L.  and 
D.  D.  VanSlyke  (1906,  1907)  showed  by  conductivity  methods  that 
in  some  cases,  at  least,  the  taking  up  of  acid  is  a  process  of  ad- 
sorption; while  Sammis  and  Hart  (Sammis,  1907a,  1907b;  Sammis 
and  Hart,  1909)  have  shown  that  the  amounts  of  the  different 


PHOSPHORUS  METABOLISM  37 

acids  required  for  coagulation  are  not  chemically  equivalent,  and 
that  the  time  required  is  affected  by  the  presence  of  other  substan- 
ces as  well  as  by  the  temperature  and  the  concentration.  All  of 
these  observations  emphasize  complication  in  the  process. 

The  papers  of  VanSlyke  and  Hart  (1902,  1905a,  1905c)  should 
be  mentioned  in  this  connection.  Other  studies  of  VanSlyke  and 
Hart  consider  the  reaction  with  acids  particularly  in  relation  to 
cottage  cheese  (1904)  and  to  the  so-called  ''mottling"  of  butter 
(1905b). 

In  the  coagulation  it  may  be  that,  in  addition  to  such  chemical 
changes  as  have  been  described,  there  is  also  concerned  a  chemico- 
physical  aggregation  of  molecules  as  suggested  by  T.  Brailsford 
Robertson  (1908)  in  the  article,  "On  the  Influence  of  Temperature 
upon  the  Solubility  of  Casein  in  Alkaline  Solutions."  In  this  con- 
nection the  nature  and  complexity  of  casein  as  a  protein  should  be 
borne  in  mind.  The  protein  molecule  always  shows  both  acid  and 
basic  qualities — is  amphoteric — indicating  that  it  yields  both  hydro- 
gen (H+)  and  hydroxyl  (OH-)  ions  on  electrolytic  dissociations  such 
as  is  conceived  to  take  place  spontaneously  in  any  water  solution. 
The  molecule  may  be  represented  by  the  formula  HXOH,  in  which 
X  represents  a  carbon  nucleus  of  variable  complexity.  Any  two 
such  molecules  may  unite,  and,  with  the  elimination  of  water,  form  a 
molecule  of  the  type  HXXOH,  according  to  the  equation : 

HX(OH+H)XOH=HXXOH+H20. 

Proteins  generally  are  made  up  by  the  union  of  simple  bodies  of 
this  type. 

According  to  Robertson's  conception,  in  a  solution  of  a  protein 
or  its  salt  the  molecules  of  the  protein  unite  with  each  other  to  a 
certain  extent,  in  this  way  forming  polymers.  The  reaction  is  re- 
versible, and  the  point  of  equilibrium  between  the  compound  and  its 
polymeric  modification  varies  under  the  influence  of  whatever  con- 
dition affects  the  concentration  of  the  protein  ions.  Addition  of 
water,  or  of  acid,  alkali  or  salt,  or  the  application  of  heat  has  such 
an  effect,  and  consequently  alters  the  relative  number  of  heavier 
molecule-complexes.  Robertson's  experiments  give  evidence  that  one 
of  the  effects  of  increase  of  temperature  on  a  solution  of  casein  is  a 
shifting  of  the  equilibrium  in  the  direction  of  the  higher  complexes. 
He  explains  coagulation  as  being  a  result  of  these  molecular  aggre- 
gates becoming  so  large  as  to  assume  the  properties  of  matter  in 
mass  and  to  become  practically  an  unstable  suspension  and  then 
a  precipitate. 


38  OHIO  EXPERIMENT  STATION:   TECHNICAL  BUL.  5 

COAGULATION  OF  CASEIN  BY  RENNET 

For  general  discussions  see  Fuld  (1902b)  and  Kastle  and  Rob- 
erts (1909). 

The  Enzyme.  Coagulation  is  brought  about  by  an  enzyme 
called  "rennin"  or  "chymosin,"  which  is  usually  obtained  from  the 
water  infusion  of  the  fourth  stomach  of  the  calf  or  sheep.  As  was 
first  noted  by  Bang  (1900c),  the  corresponding  enzyme  obtained 
from  the  gastric  juice  of  the  pig,  of  man,  and  of  some  other  animals 
differs  somewhat  from  chymosin,  and  hence  is  designated  "para- 
chymosin."  It  has  been  shown  that  this  parachymosin  action  is  to 
be  attributed  to  pepsin,  the  same  enzyme  which  has  the  proteolytic 
action  in  these  juices,  and  that  proteolytic  enzymes  generally  have 
coagulating  power  under  suitable  conditions.  Several  workers 
hold  also  that  the  distinction  between  chymosin  and  parachymosin 
is  apparent,  not  real,  and  that  chymosin  also  is  identical  with  pepsin. 
Many  articles  have  appeared,  mainly  in  Hoppe-Seyler's  Zeitschrift 
f iir  physiologische  Chemie,  on  the  two  sides  of  this  controversy. 
For  a  recent  discussion  of  evidence  for  the  identity,  see  Van  Dam 
(1912) ;  and  for  counterargument  and  evidence,  see  Rakoczy  (1913). 
Rakoczy  interprets  his  experiments,  and  harmonizes  those  of 
others,  by  the  theory  that  there  is  in  the  stomach  of  young  animals 
of  certain  species  (notably  the  calf)  an  independent  coagulating 
enzyme  accompanying  the  pepsin,  though  the  pepsin  also  has  co- 
agulating action  of  its  own  together  with  its  proteolytic  action, 
but  this  special  enzyme  is  lacking  in  the  adults  of  the 
same  species  and  in  the  young  of  other  species.  He  notes  also 
that  in  the  very  young  of  some  species  both  enzymes  are  entirely 
lacking  and  coagulation  is  due  only  to  the  acid  of  the  stomach. 

Enzymes  (or  their  zymogens)  having  rennin-like  action  are 
found  in  several  organs  of  the  higher  animals  and  in  blood  (see  Ed- 
munds, 1896),  and  they  are  said  to  occur  even  in  invertebrates,  and 
to  be  widely  diffused  in  the  plant  world.  That  of  the  pancreatic 
gland  has  been  most  studied.  (See  Roberts,  1879,  1881;  Edkins, 
1891;  Harris  and  Gow,  1892;  Halliburton  and  Brodie,  1896). 

The  Coagulum  and  Conditions  for  its  Formation.  Rennet  coag- 
ulation differs  from  coagulation  by  acids,  and  cannot  be  looked  upon 
as  a  simple  removal  of  the  base  from  a  caseinate.  The  presence  of 
soluble  calcium  salts  (or  other  alkaline  earth  salts)  seems  to  be  es- 
sential, and  the  precipitate  formed  is  not  casein  or  a  casein  salt,  but 
a  salt  of  a  slightly  different  nucleoalbumin  called  "paracasein." 
Many  writers,  following  Halliburton,  call  this  modification  produced 


PHOSPHORUS  METABOLISM  39 

by  rennin  the  "casein,"  and  that  from  which  it  is  derived,  "casein- 
ogen."  Foster  and  a  few  others  have  used  the  term  "tyrein"  for 
the  rennet  clot. 

A  number  of  investigations  have  been  made  concerning  the 
conditions  essential  or  favorable  to  formation  of  the  coagulum,  es- 
pecially with  regard  to  the  effects  of  the  degree  of  acidity  and  of 
conditions  affecting  the  amount  of  calcium  present,*  either  as  free 
soluble  salt  or  bound  to  the  casein.  (Hammarsten,  1877 ;  Schaf- 
f er,  1887 ;  Soldner,  1888 ;  Arthus  and  Pages,  1891 ;  Courant,  1891 ; 
Leze  and  Hilsont,  1894 ;  de  Jager,  1897 ;  Locke,  1897 ;  VanSlyke  and 
Hart,  1902, 1905a,  1905c;  VanDam,  1909a;  Van  Slyke  and  Bosworth, 
1912;  Bosworth,  1913;  Schryver,  1913.)  Soluble  salts  of  calcium, 
barium  and  strontium  favor  or  hasten  coagulation,  while  salts  of 
ammonium,  sodium  and  potassium  retard  or  prevent  coagulation. 

The  bulk  of  the  coagulum  from  milk  is  a  calcium  paracaseinate, 
but  it  carries  down  with  it  calcium  phosphate  and  fat,  both  of  which 
bodies  have  been  helped  to  remain  in  their  state  of  suspension  in 
milk  by  the  presence  of  the  casein  salt.  Lindet  (1912a,  1912b)  has 
concluded  that  about  one-half  of  the  phosphorus  contained  in  the 
rennet  curd  is  in  the  form  of  phosphate  of  lime  (probably  tricalcic) , 
the  other  half  being  organically  combined  phosphoric  acid. 

Hammarsten's  Interpretation  of  the  Process  of  Rennet  Coagu- 
lation. According  to  Hammarsten  (1877,  1896),  whose  view  has 
been  commonly  held,  the  distinctive  effect  of  the  ferment  is  not  pre- 
cipitation, but  is  the  transformation  of  casein  into  paracasein.  This 
is  evidenced  by  the  fact  that  if  rennet  be  allowed  to  act  on  solutions 
free  from  lime  salts  no  precipitate  occurs ;  but  there  is  an  invisible 
alteration  of  the  casein,  for  now,  even  if  the  ferment  be  destroyed 
by  boiling  the  solution,  addition  of  lime  salts  will  cause  immediate 
coagulation.  (See  also  Spiro,  1906.)  Hence  the  process  of  rennet 
coagulation  is  a  two-phase  process ;  the  first  phase  is  the  transform- 
ation of  casein  by  rennin,  the  second  is  the  visible  coagulation 
caused  by  lime  salts. 

Furthermore,  if  the  purest  casein  and  the  purest  rennin  were 
used,  Hammarsten  always  found  after  coagulation  that' the  filtrate 
contained  very  small  amounts  of  a  protein.  This  protein  he  desig- 
nated as  the  "whey  protein." 

In  accordance  with  these  observations,  Hammarsten  (1911)  ex- 
plains the  rennin  action  "as  a  cleavage  process,  in  which  the  chief 
mass  of  the  casein,  sometimes  more  than  90  percent,  is  split  off  as 
paracasein,  a  body  closely  related  to  casein,  and  in  the  presence  of 
sufficient  amounts  of  lime  salts  the  paracasein-lime  precipitates  out 
while  the  proteose-like  substance  (whey-protein)  remains  in  solu- 
tion." 


40  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

By  continued  action  of  rennin  on  paracasein  a  further  trans- 
formation has  been  found  in  several  cases  (Petry,  1906;  VanHer- 
werden,  1907;  VanDam,  1909b),  but  perhaps  due  to  a  contamination 
of  the  rennin  with  pepsin,  or  to  the  identity  of  these  two  enzymes. 
The  action  which  forms  paracasein  and  whey  protein  takes  place  in 
a  short  time  (Hammarsten,  1896;  Schmidt-Nielsen,  1906).  The 
composition  and  solubilities  of  paracasein  have  received  consider- 
able attention.  (See  Loevenhart,  1904;  Kikkoji,  1909;  VanSlyke 
and  Bosworth,  1912.)  It  is  more  readily  digested  by  pepsin-hydro- 
chloric acid  than  is  casein  (Hosl,  1910). 

Duclaux  (1884c)  and  Loevenhart  (1904)  and  others  do  not 
accept  Hammarsten's  theory;  but  to  most  workers  it  seems  prob- 
able, at  least,  that  the  action  of  the  rennin  is  to  cause  a  cleavage  of 
casein  v/ith  formation  of  paracasein.  However,  the  chemical  and 
physical  differences  observed  between  casein  and  paracasein  appear 
to  be  so  slight  that  Loevenhart  and  some  others  think  that  they  are 
only  physical,  perhaps  differences  in  the  size  of  the  colloid  or  solu- 
tion aggregates.  Loevenhart  conceives  of  a  large  part  of  the  work 
of  the  rennet  (or  of  the  acid,  in  acid  and  heat  coagulation)  as  being 
a  freeing  of  the  calcium  to  make  it  available  for  precipitation.  Some 
think  that  the  aggregates  of  paracasein  are  larger  than  those  of 
casein,  but  there  is  more  evidence  of  their  being  smaller,  which  idea 
corresponds  with  the  findings  of  Bosworth,  though  he  looks  upon 
the  change  as  a  true  cleavage. 

Bang's  Description  of  the  Precipitation.  Bang  (1911b)  studied 
the  progress  of  the  coagulation  process  by  means  of  interruptions  at 
definite  intervals.  His  observations  confirm  the  idea  that  rennin 
causes  the  formation  of  paracasein,  and  that  the  calcium  salt  serves 
only  for  the  precipitation  of  the  paracasein;  the  rennin  has  to  do 
also  with  the  mobilizing  of  lime  salts.  According  to  Bang,  before 
coagulation  occurs  paracaseins  with  constantly  greater  affinity  for 
calcium  phosphate  are  produced.  These  take  up  increasing 
amounts  of  calcium  phosphate,  until  finally  the  combination  formed 
can  no  longer  remain  in  solution. 

Bosworth's  View  of  the  Rennin  Action.  By  a  very  recent  work 
of  L.  L.  VanSlyke  and  A.  W.  Bosworth  (VanSlyke  and  Bosworth, 
1912,  1913a,  1913b,  1913c;  Bosworth  and  VanSlyke,  1913)  in  which 
ash-free  casein  and  paracasein  were  compared  as  to  their  element- 
ary composition,  and  as  to  the  salts  they  form  with  bases,  and  the 
properties  of  these  salts,  it  is  indicated  that  the  two  compounds  are 
alike  in  percentage  composition  and  in  combining  equivalent,  the 
paracasein  molecule  being  one  half  of  the  casein  molecule.  More- 
over, Bosworth  (1913)  has  shown  that,  if  the  rennin  cleavage  be 


PHOSPHORUS  METABOLISM  41 

carried  out  under  conditions  which  avoid  autohydrolysis,  no  other 
protein  is  formed ;  also  that,  if  the  calcium  casemate  present  be  one 
containing  four  equivalents  of  calcium,  the  paracaseinate  does  not 
precipitate,  save  in  the  presence  of  a  soluble  calcium  salt,  while,  if 
the  calcium  casemate  be  one  of  two  equivalents  of  base,  rennin  does 
cause  immediate  coagulation.  Bosworth  concludes  that  the  rennin 
action  is  a  cleavage  (probably  hydrolytic)  of  a  molecule  of  caseinate 
into  two  molecules  of  paracaseinate,  the  coagulation  being  a  second- 
ary effect  due  to  a  change  in  solubilities,  dicalcium  paracaseinate 
being  soluble  in  pure  water  but  not  in  water  containing  more  than  a 
trace  of  calcium  salt,  and  the  monocalcium  caseinate  being  insolu- 
ble in  water.  The  alkali  paracaseinates,  as  well  as  caseinates,  are 
soluble.  This  explanation  seems  to  promise  to  harmonize  the  ob- 
servations with  regard  to  acidity  and  the  effects  of  the  presence  of 
soluble  salts. 

The  investigations  of  these  authors  and  of  Hart  with  regard 
to  the  changes  which  the  paracasein,  the  calcium  and  the  phospho- 
rus undergo  during  the  ripening  of  cheese  (VanSlyke  and  Hart,1902, 
1905a,  1905c;  VanSlyke  and  Bosworth,  1907,  1913d;  Bosworth, 
1907)  contributed  toward  this  interpretation. 

Mellanby  (1912)  holds  an  altogether  different  idea.  He  rep- 
resents the  coagulation  of  milk  as  due  to  an  absorption  of  a  proteo- 
lytic enzyme  (rennin  is  considered  to  be  identical  with  pepsin)  by 
the  casein,  and  the  precipitation  of  this  complex  by  calcium  ions,  or 
other  such  ions,  the  calcium  being  supposed  to  act  by  reason  of  its 
electric  charge  without  entering  into  chemical  union. 

Schryver  (1913)  has  a  theory  that  the  action  of  the  enzyme  is 
to  free  the  surface  of  the  complex  colloidal  molecules  from  adsorbed 
compounds  which  interfere  with  aggregation. 

HYDROLYTIC  CLEAVAGE  OF  CASEIN 

Since  casein  is  a  protein,  it  undergoes  hydrolysis  when  boiled 
with  mineral  acids  or  alkalis,  or  when  warmed  with  the  digestive 
proteolytic  enzymes.  Under  the  influence  of  any  of  these  agents 
the  elements  of  the  water  molecule  enter  into  the  molecule  of  the 
protein,  and  the  protein  molecule  splits  into  simpler  units.  Aside 
from  the  phosphoric  acid,  the  nature  of  the  intermediate  and  final 
products,  as  far  as  known,  is  the  same  for  casein  as  for  other  pro- 
teins.     The  processes  are  complex  and  but  partially  understood. 

The  latest  complete  analysis  of  casein  which  we  have  seen  is 
that  of  T.  B.  Osborne  and  H.  H.  Guest  (1911).  From  their  own  de- 
terminations and  from  all  data  previously  published  they  collected 
a  table  of  the  highest  values  which  had  been  found  up  to  that  time 


42 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


by  methods  which  were  considered  reliable.  It  is  to  be  noted  that 
there  is  still  about  a  third  of  the  molecule  unaccounted  for,  owing  to 
faults  in  the  methods  or  difficulties  in  manipulation.  We  give  their 
table  below  together  with  the  only  other  such  analyses  we  have 
found  which  are  at  all  extensive.  (Abderhalden,  1905;  Osborne 
and  Guest,  1911 ;  Abderhalden  and  Schittenhelm,  1906b ;  Abderhal- 
den and  Langstein,  1910.  See  also  Abderhalden  and  Funk,  1907, 
and  those  referred  to  in  the  table.) 


HYDROLYTIC  CLEAVAGE  PRODUCTS  OF  CASEIN— Percent 


Casein    from 

jow's    milk 

From  goat's  milk 

From  human  milk 

Cleavage 

products 

Formulae 

Tahle    of    Ab- 
derhalden, 

Table    of     Os- 
borne     and 

Analysis     by    Ab- 
derhalden  and 
Schittenhelm, 
1906b     ' 

Analysis    by    Ab- 
derhalden and  - 

1905 

Guest,   1911 

Langstein,       1910 

Glycocoll 

C-,HBN02 

0.00 

0.00 

Alanine 

C3H7N02 

0.9 

1.50  1 

1.5 

1.2 

Valine 

C5HuN02 

1.0  * 

7.20  x 

1.3 

Leucine 

CoHisNOa 

10.5 

9.35  2 

7.4 

8.8 

Proline 

C5HeN02 

3.1  f 

6.70  3 

4.62 

2.85 

Phenylalanine 

CHnNOs 

3.2 

3.20  4 

2.75 

2.8 

Glutaminic 

CbHbNO* 

10.7 

15.55   1 

11.25 

10.95 

acid 

Aspartic  acid 

C4H7NO4 

1.2 

1.39  1 

1.1 

1.0 

Cystine 

CeH1204N2S2 

0.0655 

1 

Serine 

C3H7N03 

0.23    6 

0.50  8 

Tyrosine 

CoHuNOs 

4.5 

4.50  7 

4.95 

4.58 

Oxyproline 

C5H3N03 

0.25§8 

0.23  8 

Histidine 

C<,H9N302 

2.59   9 

2.50  10 

Arginine 

C8H14N402 

4.84  9 

3.81  10 

Lysine 

C6H14N202 

5.80  9 

5.95  10 

Tryptophane 

CxiH^NaOo 

1.5 

1.50  i 

Diaminotri- 

oxydodecanic 

acidj 

Cn>H26N205 

0.75  u 

0.75  "■ 

Present 

Ammonia 

NH3 

1.61  u 

Sulphur 

s 

0.76  13 

Phosphorus 

P 

0.85  13 

Sum 

51.13 

67.85 

(*)  Amino-valerianic  acid.  (f)  Pyrrolidine-carboxylic  acid.  (§)  Oxy- a  -pyrrolidine- 
carboxylic   acid.      (})      Sometimes   called    "caseinic   acid." 

Authors  to  whom  credit  is  given:  (1)  Osborne  and  Guest.  (2)  Levene  and  VanSlyke, 
original  has  "leucine+isoleucine."  (3)  VanSlyke.  (4)  Abderhalden.  (5)  K.  A.  H. 
Morner.  (6)  Fischer,  original  has  0.5  percent.  (7)  Reach.  (8)  Fischer,  original  has 
0.23  percent.  (9)  E.  Hart.  (10)  Osborne,  Leavenworth  and  Brautlecht.  (11)  Fischer 
and  Abderhalden.        (12)  Osborne  and  Harris.        (13)  Hammarsten. 

Folin  and  Denis  (1912)  have  recently  reported  the  determina- 
tion of  tyrosine  by  a  new  colorimetric  method.  They  find  6.5  per- 
cent in  casein.  E.  Fischer  and  Abderhalden  (1903)  by  enzyme  di- 
gestions found  a-pyrrolidine  carbonic  acid. 


PHOSPHORUS  METABOLISM  43 

With  regard  to  the  relation  of  phosphorus  to  the  casein  mole- 
cule, Osborne  and  Guest  say  in  their  discussion:  "The  presence  of 
phosphorus  in  casein  raises  the  question  as  to  whether  it  is  a  con- 
stituent of  the  protein  molecule  or  of  some  non-protein  group  united 
with  a  protein  group  as  is  a  haematin  with  globin  in  haemoglobin, 
or  nucleic  acid  with  protein  in  the  nucleins.  Such  data  as  are  avail- 
able indicate  that  the  union  is  other  than  a  salt-like  combination  of 
a  phosphorus-containing  acid  with  a  protein  base."  Calculations 
from  the  amount  of  the  casein  and  of  its  nitrogen  unaccounted  for 
by  the  above  analyses  "give  no  evidence  that  casein  differs  in  con- 
stitution to  any  marked  degree  from  other  proteins  which  contain 
no  phosphorus.       It  might  be  supposed  that  the  phosphorus  of 

casein  was  a  part  of  some  organic  radical If  this  is,  in  fact, 

so  the  organic  radical  must  be  one  which  contains  nitrogen  in  ap- 
proximately the  same  proportion  as  the  mono-amino-acids." 

DIFFERENCES  IN  THE  CASEIN  FROM  DIFFERENT  KINDS   OF  MILK 

Consistency  of  the  Coagulum.  There  is  a  marked  difference  in 
the  appearance  and  the  digestibility  of  the  coagulum  formed  by 
acids  or  rennet  in  cow's  milk  and  human  milk.  The  milk  of  the 
mare,  and  the  ass  (Langgaard,  1875;  Ellenberger,  1899;  Storch, 
1902;  Zaitschek,  1904;  von  Szontagh,  1905)  are  said  to  resemble 
human  milk  in  this  respect  and,  according  to  Arthus  and  Pages 
(1891),  the  milk  of  the  dog  also  belongs  in  this  class.  The  cow's 
milk  gives  a  much  tougher  and  more  compact  coagulum,  in  coarser 
flakes,  than  the  small,  jelly-like  flakes  from  human  milk.  This 
difference,  however,  may  not  be  due  to  any  chemical  difference  in 
the  casein  itself.  Certain  observations  indicate  that  it  might  be 
due  to  the  greater  proportionate  amount  of  fat  in  the  human  milk 
(Hempel,  1894),  or  of  salts  in  cow's  milk  (Dogiel,  1885),  or  to  the 
acidity  of  cow's  milk  (Courant,  1891).  Soxhlet  (1893)  says  that 
dilution  of  cow's  milk  reduces  the  size  of  the  flakes.  Biedert  and 
Schroter  (Biedert,  1887 ;  Schroter,  1887)  notice  great  differences  in 
the  amount  precipitable  with  MgS04.  Ass's  milk  is  not  easily  co- 
agulated by  acids  and  its  casein  not  easily  salted  out  by  salts 
(Storch,  1902),  and  the  coagulum  is  fine. 

Completeness  of  Peptic  Digestion.  Early  investigators  report 
that  peptic  digestion  of  cow's  casein  leaves  an  insoluble,  undigested 
nuclein  (pseudonuclein)  but  casein  from  human  milk  does  not  (von 
Szontagh,  1892,  1894;  Willdenow,  1893;  von  Moraczewski,  1895a; 
Alexander,  1898) ;  but  later  work  seems  to  prove  that  this  differ- 
ence is  merely  one  of  the  ease  of  digestion  of  the  two  pseudonu- 
cleins.  (Wroblewski,  1894a,  1894b ;  Sebelien,  1894,  1895 ;  Salkow- 
ski,  1893a,  1893b,  1896a;  Salkowski  and  Hahn,  1894-5;  Kobrak, 


44  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1900;  Rotondi,  1902).  Salkowski  says  that  the  casein  will  be  com- 
pletely digested  if  the  ratio  between  casein  and  digesting  liquid  is 
1:500  and  the  casein  has  previously  been  dissolved  so  that  the  in- 
fluence of  hard,  dried  particles  of  casein  is  excluded.  About  1  per- 
cent remains  undissolved  if  the  ratio  of  casein  to  solvent  is  1 :250. 
Zaitschek  and  von  Szontagh  (Zaitschek,  1904;  von  Szontagh,  1905) 
found  woman's,  ass's  and  mare's  milk  completely  digested  in  72 
hours  at  38°C,  while  only  8,  14  and  15  percent,  respectively,  of  the 
casein  of  cow's,  buffalo's  and  goat's  milk  is  rendered  soluble  under 
the  same  conditions.  Long  (1906a,  1907a)  finds  the  caseins  of 
goat's  and  cow's  milk  much  alike,  but  the  former  has  a  somewhat 
higher  equivalent  weight  than  the  latter,  and  it  is  more  slowly  acted 
on  by  pepsin-hydrochloric  acid,  and  leaves  more  pseudonuclein. 

Elementary  Composition.  By  reference  to  the  table  on  p.  35  it 
will  be  seen  that  there  is  not  much  difference  in  the  elementary  com- 
position of  caseins  from  various  sources  except  as  to  the  phosphorus 
content.  Probably  the  analyses  of  Langstein  and  Edelstein  (1910) 
are  the  best  we  have  on  human  casein.  Then,  as  compared  with 
about  0.85  percent  phosphorus  in  cow's  casein,  we  find  only  about 
0.24  percent  in  human  casein  and  as  high  as  1.04  percent  in  ass's 
casein. 

Products  of  Complete  Hydrolysis.  The  analyses  of  Abderhal- 
den  and  his  associates  (see  p.  42)  show,  so  far  as  they  go,  no  marked 
differences  between  the  cleavage  products  of  the  caseins  from  cow's, 
goat's  and  woman's  milk,  considering  the  lack  of  close  agreement  of 
all  such  determinations.  On  the  other  hand,  they  by  no  means  ex- 
clude the  possibility  of  significant  differences  of  this  kind,  as  ac- 
counting for  the  variations  in  elementary  composition  and  in  chem- 
ical behavior. 

Other  Comparisons.  In  the  other  ways  in  which  caseins  have 
been  compared — as  to  solubilities,  reactions  with  chemicals,  repeat- 
ed solution  and  reprecipitation,  pancreatic  digestion  and  nutritive 
value — there  seems  to  be  enough  similarity  observed  to  indicate  that 
all  the  caseins  are  alike  in  general  nature,  but  unlike  in  some  chemi- 
cal respects  which  are  not  yet  determined.  J.  Bauer  and  St.  Engel 
(1911)  concluded  that  the  caseins  from  cow  and  human  milk  are 
alike. 

THE    PHOSPHORUS    OF    CASEIN 

What  is  to  be  said  as  to  the  form  in  which  phosphorus  is  present 
in  casein  ?  The  quotation  from  Osborne  and  Guest  shows  that  practi- 
cally nothing  is  known  as  to  the  nature  of  that  within  the  protein 
itself,  not  even  whether  it  be  present  as  a  phosphoric  acid  radical 


PHOSPHORUS  METABOLISM  45 

or  as  a  nitrogenous,  organic  complex.  Aside  from  the  phosphorus 
within  the  protein,  casein  is  also  practically  always  intimately  as- 
sociated with  calcium  phosphate. 

Perhaps  the  synthetic  work  of  Neuberg  and  Pollak  (1910b, 
1910d)  throws  some  light  on  the  nature  of  the  combination  of  phos- 
phorus in  the  phosphoprotein  molecule,  as  well  as  that  in  the  nuclein 
molecule.  At  least,  they  have  succeeded  in  bringing  about  a  union 
of  simple  proteins  (lactalbumin  and  blood  globulin)  with  the  phos- 
phorus of  phosphorus  oxychloride.  A  substance  obtained  in  this 
way  had  an  elementary  composition  much  like  that  of  casein  (but 
with  somewhat  higher  phosphorus  content)  and  yielded  to  pancre- 
atic and  peptic  digestion  in  just  about  the  same  way  as  casein.  The 
authors  looked  upon  this  compound  as  a  substituted  phosphaminic 
acid.  With  the  simple  amino  acids  or  their  esters  they  were  able 
also  to  cause  organic  union  of  the  phosphorus,  but  the  organic  phos- 
phorus compounds  were  not  isolated. 

OTHER  PHOSPHOPROTEINS 
OVOVITELLIN 

Ovovitellin  is  the  lecithalbumin  found  in  the  yolk  of  hen's  eggs. 
It  is  a  combination  of  protein  with  lecithin,  the  lecithin  not  being 
removable  by  ether.  According  to^  T.  B.  Osborne  and  Campbell 
(1900b)  the  body  which  has  been  studied  under  this  name  is  a  mix- 
ture of  various  vitellin-lecithin  combinations,  containing  from  15  to 
30  percent  of  lecithin.  The  protein,  moreover,  when  freed  from 
lecithin  still  contains  phosphorus.  Osborne  and  Campbell  give  the 
composition  of  this  protein  (which  they  call  "nucleovitellin")  : 
C=51.24;  H=7.16;  N=16.38;  S=1.04;  P=0. 94;  0=23.24  percent. 
Hammarsten  (1911)  tells  us  that  Gross  (Zur  Kenntniss  des  Ovovi- 
tellin, Inaug.  Diss.,  Strassburg,  1899)  gave  the  analysis  of  vitellin 
prepared  by  (NH4)2S04  precipitation  as  follows :  C=48.01 ;  H=6.35  ; 
N=14.91— 16.97;  P=0.32— 0.35;  S=0.88  percent;  and  we  have 
from  T.  B.  Osborne  (1902)  the  formula  C671H1112N182S5P40,.7,  with 
0.82  percent  P. 

Plimmer  (1908)  found  0.65-1.14  percent  P,  with  a  mean  of  0.99 
percent  in  five  preparations  examined.  Plimmer  found  with  the 
vitellin  in  egg-yolk  another  protein  having  a  lower  phosphorus  con- 
tent (0.10-0.85  percent  P,  with  a  mean  of  0.35  percent  in  five  prepar- 
ations), which  he  suggests  may  be  vitellin  without  its  phosphorus- 
containing  portions.  He  gave  the  new  substance  the  name  "live- 
tin." 

Abderhalden  and  Hunter  (1906)  and  T.  B.  Osborne  and  D.  B. 
Jones  (1909)  report  the  following  cleavage  products: 


46  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

CLEAVAGE  PROUCTS  OF  VITELLIN  OF  HEN'S  EGG— Percent 


Acid 


Glycocoll     

Alanine      

Amino-valerianic  acid   (Valine) 

Leucine     

Aspartic  acid   

Glutaminie  acid 

Phenylalanine     , 

Proline    

Serine     

Tyrosine 

Cystine    

Histidine    

Arginine   

Lysine     • 

Ammonia     , 

Tryptophane     

Phosphorus     , 

Total     


Abderhalden  and  Hunter 
(1906) 


1.1 

Present 

2.4 
11.0 

0.5 
12.2 

2.8 

3.3 

1 

i.e 

Not  determined 


Osborne  and  Jones   (1909) 


Not   complete 


0.00 
0.75 
1.87 
9.87 
2.13 
12.95 
2.54 
4.18 
3 

3.37 
Not  determined 
1.90 
7.46 
4.81 
1.25 
Present 
0.94 


Folin  and  Denis  (1912)  found  in  ovovitellin  5.2  percent  tyrosin. 

The  study  of  Osborne  and  Campbell  already  cited  includes  ex- 
amination of  the  paranuclein  resulting-  from  peptic  digestion.  By  re- 
peated digestions  they  obtained  substances  containing  3.29,  2.52  and 
4.19  percent  P.  They  interpret  their  analyses  as  showing  that  this 
paranuclein  and  the  paranucleoprotein  from  which  it  was  derived 
"are  both  compounds  of  one  and  the  same  proteid  body  [which  is 
free  from  phosphorus,  and  which  these  authors  call  "vitelline"] 
with  a  phosphoric  acid,  possiblyH3P04,  HsP209,  or  some  simple 
organo-phosphoric  acid."  For  the  paranucleic  acid  prepared  by  Lev- 
ene  and  Alsberg  (1901)  the  phosphorus  content  is  given  as  9.88 
percent  P. 

Bunge  (1885a)  made  a  study  of  the  paranuclein  derived  from 
peptic  digestion  of  the  egg-yolk  as  a  whole.  In  this  body  he  found 
5.19  percent  P,  Hugounenq  and  Morel  (1905a,  1905b)  later  finding 
8.7  per  cent  P.  This  body  Bunge  looked  upon  as  that  from  which 
haemoglobin  is  made  for  the  young  organism,  and  he  gave  it  the 
name  "haematogen."  The  significance  of  the  iron  content  is 
emphasized. 

Finally,  it  is  evident  that  much  more  study  may  well  be  given 
to  the  constitution  and  metabolic  possibilities  of  ovovitellin,  which 
is  so  significant  for  the  development  of  the  young  bird,  and  import- 
ant as  a  food  for  man.      Phosphorus  is  present  in  a  lecithin  portion 


PHOSPHORUS  METABOLISM 


47 


and  in  a  protein  p.ortion,  and  probably  the  two  are  chemically  com- 
bined. In  the  lecithin  fraction  it  is  glycerylphosphate,  and  in  the 
protein  fraction  some  undetermined  radical,  very  likely  a  simple 
phosphoric  acid  or  a  phospho-organic  acid. 

ICHTHULIN 

The  compound  in  the  eggs  of  fish  corresponding  to  the  ovovi- 
tellin of  hen's  eggs  has  been  given  the  name  "ichthulin."  The  two 
were  early  studied  together  and  their  similarities  in  chemical  nature 
as  well  as  biological  significance  were  pointed  out.  (See  Gobley 
1850a ;  Valenciennes  and  Fremy,  1854 ;  Diaconow,  1867a ;  G.  Walter, 
1891;  Levene,  1901a;  Hammarsten,  1905a;  Plimmer  and  Scott, 
1908;  Gobley  quotes  other  authors  as  far  back  as  1817.)  Such  a 
'body  has  been  isolated  and  studied  from  the  eggs  of  carp,  sturgeon, 
cod  and  perch.  Levene  (1901a)  formed  the  paranuclein  by  peptic 
digestion,  and  reports  the  analysis  of  the  paranucleic  acid,  which 
corresponds  with  that  of  the  product  from  ovovitellin.  This  acid 
(not  entirely  free  from  protein)  showed  10.34  percent  P.  We  give 
below  Levene's  table  of  the  analyses  of  ichthulin  made  by  different 
workers,  and  add  values  taken  from  Hammarsten's  (1905a)  later 
report. 

ELEMENTARY  ANALYSIS  OF  ICHTHULIN  FROM  FISH  EGGS— Percent 


Author 

Kind  of  fish 

C 

H 

N 

S 

P 

Pe 

-  0 

Fremy 

Salmon 

52.5- 
53.3 

8.82 

15.2 

1.00 

0.6 

22.7 

Gobley 

Carp 

52.6 

7.74 

15.5 

0.90 

0.37 

23.24 

Walter 

Carp 

53.52 

7.6 

15.63 

0.41 

0.43 

0.10 

22.19 

Levene 

Cod 

52.44 

7.45 

15.96 

0.92 

0.65 

22.58 

Hammarsten 

Perch 

14.81 

1.111 

0.743 

OTHER  PHOSPHOPROTEINS  OF  ANIMAL  ORIGIN 

Hammarsten  (1885b)  reports  obtaining  a  nucleoalbumin  from 
the  albuminous  gland  of  a  snail.  This  body  is  spoken  of  as 
"helicoproteid  from  the  name  of  the  snail  (Helix  pomatia) .  Lieber- 
mann  (1891a,  1891b,  1893a,  1893b)  isolated  what  he  called  lecithal- 
bumins  from  the  mucous  membrane  of  the  stomach  (hog),  from 
kidney  (sheep),  liver  (lamb),  lungs  and  spleen.  He  called  especial 
attention  to  the  observations  that  these  substances  are  strongly 
acid,  and  readily  form  union  with  alkalis  or  other  basic  bodies,  that 
they  are  rendered  still  more  acid  by  C02 ;  then  by  soda  solutions  are 


48  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

rendered  strongly  alkaline,  and  afterward  can  again  be  made  acid  by 
C02.  He  suggests  that  in  the  stomach  walls  they  may  thus  serve  in 
the  production  of  hydrochloric  acid  from  sodium  chloride  by  taking 
up  the  sodium  temporarily  and  gradually  giving  it  up  to  the  C02  of 
the  blood.  He  also  found  that  the  alkaline  solutions  of  Na,HP04,  or  of 
sodium  urate  carrying  a  slight  excess  of  soda,  when  passed  through 
these  lecithalbumins  as  a  filter,  show  a  strongly  acid  nitrate,  and 
leave  a  strongly  alkaline  residue  on  the  filter.  This  he  suggests 
may  indicate  the  process  by  which  an  acid  urine  may  result  in  the 
kidney  from  filtration  from  alkaline  blood. 

Lonnberg  (1890)  looking  for  mucin  in  the  cortical  and  medul- 
lary substance  of  the  kidney  and  the  mucous  membrane  of  the  urin- 
ary bladder,  decided  that  the  bodies  he  isolated  were  not  true 
mucins,  but  were  nucleoalbumins.  Malengreau  (1900)  found  two 
nucleoalbumins  in  the  thymus. 

Plimmer  and  Kaja  (1909) ;  also  Plimmer  and  Scott  (1908)  re- 
port phosphoproteins  in  the  pancreas  and  pancreatic  juice  of  the 
dog,  in  the  salivary  glands  of  sheep  and  in  the  eggs  in  the  ovary 
of  the  frog.  They  did  not  find  them  in  the  testes  of  the  ox  or  of 
codfish,  nor  in  thymus. 

PHOSPHOPROTEINS  OF  VEGETABLE  ORIGIN 

From  time  to  time  proteins  containing  phosphorus  have  been 
isolated  from  plant  bodies,  and  some  of  these  so  closely  resemble  the 
animal  phosphoproteins  in  their  properties  that  Liebig  gave  them 
the  name  of  vegetable  casein;  this  name,  however,  was  generally 
given  up  when,  later,  Weyl  showed  that  none  of  the  substances  most 
resembling  casein  are  present  in  the  natural  bodies,  but  that  there 
are  there  such  as  may  be  called  plant  vitellins — phyto-vitellins.  All 
of  the  investigations  of  these  compounds  are  thrown  somewhat  in 
doubt  because  of  the  presence  of  considerable  amounts  of  salts  such 
as  potassium,  calcium,  or  magnesium  phosphates,  or  of  alkalis  in 
combination  with  organic  acids,  which  are  very  difficult  of  removal. 
Hammarsten  (1911,  p.  105)  saj^s:  "It  is  not  clear  whether  the  phos- 
phorized  plant  proteids  contain  their  phosphorus  as  impurities  or 
whether  they  are  the  same  as  the  animal  phosphoproteins." 

The  vegetable  vitellins  that  have  been  most  thoroughly  inves- 
tigated are,  according  to  Gustav  Mann,  (Chemistry  of  the  Proteids, 
1906  pp.  374,  375;  23  references  on  this  subject),  the  gluten-casein 
of  wheat ;  similar  substances  from  rye,  maize,  spelt  and  barley ;  leg- 
umins  from  peas,  vetches,  beans,  lentils,  etc.  (T.  B.  Osborne  says 
that  these  are  globulins) ;  conglutins  (so  named  by  Ritthausen)  of 
lupines,  almonds,  nuts,  etc.  Several  of  these  have  been  analyzed 
by  Ritthausen  and  by  Osborne. 


PHOSPHORUS  METABOLISM  49 

We  have  not  found  report  of  any  recent  work  on  any  of  the 
compounds  of  this  group. 

PHOSPHOCARNIC  ACID 

Phosphocarnic  acid  is  a  complex  phospho-nitrogenous  com- 
pound the  nature  of  which  seems  never  to  have  been  fully  deter- 
mined, the  substance  never  having  been  isolated  free,  but  only  in 
its  iron  compound,  carniferrine,  or  mixed  with  decomposition  prod- 
ucts. It  was  first  described  by  Siegfried  in  1894  (see  Siegfried 
1894,  1895,  1896)  who  obtained  it  at  first  from  prepared  meat  ex- 
tracts and  later  directly  from  muscle.  It  is  perhaps  the  most  im- 
portant organic  phosphorus  compound  among  the  meat  extractives. 
The  properties  make  it  well  fitted  to  serve  as  a  carrier  of  phosphoric 
acid,  iron,  lime  and  magnesia  in  the  body  fluids,  since  it  readily 
unites  with  the  basic  elements  and  forms  with  them  compounds 
soluble  in  either  neutral,  weakly  acid  or  weakly  alkaline  solutions. 
It  probably  has  a  similar  significance  in  milk,  of  which  also  it  is  a 
constant  constituent. 

The  cleavage  products  which  Siegfried  obtained  from  phospho- 
carnic acid  are  carnic  acid  (which  seems  to  be  identical  with,  or 
closely  related  to,  the  antipeptone  of  Kuhne) ,  phosphoric  acid,  car- 
bon dioxide,  a  carbohydrate  group  which  reduces  Fehling's  solution, 
succinic  acid  and  paralactic  acid.      (See  also  T.  R.  Kruger,  1896.) 

Since  carnic  acid  is  a  peptone,  phosphocarnic  acid  differs  from 
nucleins  primarily  in  that  on  hydrolytic  cleavage  it  yields  a  peptone 
direct  instead  of  a  protein.  For  such  compounds  Siegfried  pro- 
posed the  name  "nucleon,"  and  he  designated  phosphocarnic  acid  as 
muscle-nucleon." 

The  nucleon  obtained  from  milk  showed  nearly  the  same  com- 
position as  that  from  muscle,  but  yielded  decomposition  products  of 
slightly  different  properties ;  the  lactic  acid  obtained  was  not  para- 
lactic but  was  fermentation  lactic  acid,  and  in  place  of  carnic  acid 
there  was  obtained  a  modification  designated  as  orylic  acid. 

T.  R.  Kriiger  (1899)  states  that  milk  nucleon  is  not  precipitated 
by  ammonium  sulphate,  while  the  muscle  nucleon  is  so  precipi- 
tated, and  that  such  precipitation  alters  the  proportion  of  N  to  P. 
Siegfried  (1899)  pointed  out  at  the  same  time  that  the  precipitates 
which  he  and  others  were  obtaining  from  muscle  were  not  uniform 
in  the  ratio  of  nitrogen  to  phosphorus,  which  might  be  due  to  the  ex- 
istence in  muscle  of  nucleons  of  different  composition.  That  would 
not  be  the  only  possible  explanation  of  this  inconsistency,  however, 
for  it  is  possible  that  differences  in  the  conditions  of  precipitation 


50  OHIO  EXPERIMENT  STATION:  TECHNICAL  BTJL.  5 

brought  down  a  mixture  of  substances,  or  even  caused  some  slight 
decomposition  of  the  nucleon.  In  Siegfried's  analyses  of  carnifer- 
rine  from  muscle  reported  in  1896  the  nitrogen  content  ranged  from 
5.45  to  6.03  percent,  the  mean  of  six  determinations  being  5.65  per- 
cent. The  phosphorus  content  found  in  four  cases  ranged  from 
1.84  to  2.59  percent  (P),  the  mean  being  2.12  percent.  Using  the 
mean  value  for  nitrogen,  and  relating  it  to  the  lowest  and  highest 
values  for  phosphorus,  gives  the  ratio  N  :P  as  3.07  and  2.18  respect- 
ively. The  ratio  N:P  found  at  this  time  in  a  preparation  from 
muscle  of  a  new  born  calf  was  as  1:1,  and  Kriiger  had  found  a  ratio 
of  1 :1  during  his  work  with  muscle  from  a  steer.  Macleod  (1899), 
in  the  same  laboratory,  had  precipitated  carniferrine  from  the  ex- 
tracts of  muscle  from  a  number  of  dogs,  and  had  found  that  they 
varied  in  this  respect.  The1  analyses  he  gives  of  nine  such  extracts 
show  these  values  for  N:P,— 4.3,  1.6,  1.8,  1.5,  1.7,  1.5,  1.3,  2.0,  3-.7. 

This  all  gives  sufficient  evidence  that  we  have  not  a  uniform 
product,  and  that  if  phosphocarnic  acid  is  a  chemical  unit  it  under- 
goes decomposition  in  the  processes  used  in  isolation. 

The  animal  substances  in  which  phosphocarnic  acid  has  been 
found  (or,  rather,  from  the  water  extracts  of  which  its  iron  complex 
has  been  precipitated  after  removal  of  proteins  and  phosphates)  are 
as  follows: 

By  Siegfried  (1896) :  Liebig's  and  Kemmerich's  meat  extract, 
beef  muscle,  dog's  liver  and  heart,  and  cow's  milk. 

By  Balke  and  Ide  (1896) :  Kemmerich's  meat  extract,  the  heart, 
liver  and  kidney  of  horse  and  of  dog. 

By  Wittmaack  (1897) :  The  milk  of  cow,  woman  and  goat. 

By  Panella :  The  striated  muscle  fibre  of  dogs  and  rabbits  (Pan- 
ella,  1902a,  1903b) ;  brain  of  dog,  rabbit  and  calf  (Ditto,  1902b, 
1903a) ;  blood  of  dog,  rabbit  and  calf  (Ditto,  1902c,  1903c) ;  both 
white  and  red  muscle  of  rabbit  (Ditto,  1903e,  1903f ) ;  testicular  sub- 
stance of  ass  and  horse  (Ditto,  1903d) ;  both  white  and  gray  matter 
of  brain  (Ditto,  1903g,  1903h) ;  testicular  substance  of  horse  (Ditto, 
1903i,  1904a)  ;  non-striated  muscle,  in  larger  amount  than  in  striat- 
ed (Ditto,  1904b) ;  spleen  of  cattle,  horses,  sheep,  swine  and  dogs 
(Ditto,  1904c) ;  the  amount  in  the  brain  of  the  dog  falls  off  during 
fast  (Ditto,  1906a). 

In  1906  Panella  (1906b)  published  a  paper  which  seems  to 
throw  doubt  on  the  numerical  data  of  previous  work  by  showing 
that  the  values  found  for  muscle  are  too  high  if  less  than  20  gm.  of 
substance  are  taken  as  the  sample.      Using  50gm.  or  more  as  sample 


PHOSPHORUS  METABOLISM  51 

yielded  nearly  constant  results  of  about  0.01  percent  nucleon  in 
fresh  muscle,  or  0.02-0.03  percent  in  dry.  These  values  are  much 
lower  than  those  which  were  earlier  reported. 

PHYTIN 

Introduction.  The  literature  of  the  chemical  study  of  phytin 
has  recently  been  reviewed  by  A.  R.  Rose  (1912b)  with  especial 
reference  to  its  significance  in  plant  economy.  This  review  is  pub- 
lished in  the  Biochemical  Bulletin,  and  is  the  most  complete  discus- 
sion to  which  we  can  refer  the  reader. 

Phytin  is  the  name  generally  applied  to  the  alkali  and  alkaline 
earth  salts  of  an  organic  phosphoric  acid  which  is  found  ex- 
tensively in  the  vegetable  world,  and  which  by  most  investigators 
is  supposed  to  contain  inosite  combined  in  some  way  with  phosphor- 
ic acid.  It  appears  now  probable  that  the  natural  products  are 
not  all  identical,  and  distinction  must  be  made  between  the  salts  of 
phytic  acid  and  of  other  phyto-phosphoric  acids.  Rose  calls  the 
acid  "inosite-phosphoric  acid." 

Discovery.  Rose  tells  us  that  the  aleurone  grains,  in  which 
this  compound  is  found  in  seeds,  were  first  discovered  microscopi- 
cally by  Hartig  in  1854,  and  that  the  particular  P-bearing  spheroid- 
al bodies  in  these  grains  were  isolated  by  Pfeffer  in  1872,  and  named 
by  him  "globoid."  Chemical  study  began  with  Palladin  in  1893, 
who  obtained  it  from  Senapis  niger.  He  showed  the  compound  to  be 
rich  in  phosphorus,  and  to  contain  magnesium  and  calcium  but  no 
nitrogen ;  also  that  it  was  non-reducing  and  yielded  no  reducing  sub- 
stance on  acid  hydrolysis.  Palladin's  work  was  followed  up  by 
Schulze  and  Winterstein  (1896)  and  Winterstein  (1897)  who  identi- 
fied the  body  with  the  globoid  of  Pfeffer.  The  name  "inosite-phos- 
phoric acid"  was  proposed  by  these  investigators  in  accordance 
with  their  finding  that  the  magnesium  salt  on  being  digested  for  30 
hours  with  cone,  hydrochloric  acid  at  130-140°  yielded  inosite  and 
phosphoric  acid.  Posternak  studied  the  substance  extensively 
(Posternak,  1900,  1903a,  1903b,  1903c,  1903d,  1905 ;  and  Bull.  Soc. 
chim.  33  (1904),  116).  He  considered  that  the  inosite  was  not  pre- 
existent  in  the  molecule,  but  that  the  compound  was  an  anhydro- 
oxymethylene-diphosphoric  acid,  and  he  gave  it  the  name  "phytin," 
which  has  been  retained. 

Occurrence.  Phytin  is  found  most  frequently  in  the  aleurone 
of  seeds  or  in  other  parts  of  plants  which  serve  for  storage,  as  some 
roots  and  tubers.  Of  investigations  as  to  the  amount  of  phosphorus 
present  in  this  form  we  mention  Schulze  and  Winterstein  (1896), 


52 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


Posternak  (1903a,  1903b),  Hart  and  Andrews  (1903),  Patten  and 
Hart  (1904),  Suzuki  and  Yoshimura  (1907),  Suzuki,  Yoshimura  and 
Takaishi  (1907),  Vorbrodt  (1910),  Rising  (1910),  Hart  and  Totting- 
ham  (1910),  and  Bernardini  (1912). 

ANALYTICAL  DATA  ON  THE  OCCURRENCE  OF  ORGANIC  PHOSPHATE 
PHOSPHORUS,  "PHYTIN"— Percent 


Substance  analyzed 


Phosphorus 
in  the  form 
of  organic 
phosphate 


Fraction  of 
total  phos- 
phorus in 
this  form 


Black  mustard  (Si/tapis  nigra)  (1) 

Red  fir 

Spruce  seed  {Picea  cxcctsa)  (2) 

Pine  seed  (Pinus  ccmbra)  (2) 

Hempseed  (cortex  removed) 

Hempseed    (  Cannabis  sativa)  (2) 

Sunflower  seed  (cortex  removed) 

Pea 

Pea,  yellow 

White  kidney  bean 

Bean,  brown 

Lentils 

Lentils  {Lens  escuZentd)  (2) 

Windsor  bean  {.Viciaf aba  minor)  (2) 

Wheat  bran 

Wheat  bran  (3) 

Wheat  grain  (4) 

Wheat  grain   (Triticum  sativum)  (2) 

Graham  flour 

Rice  bran  (3) 

Rice  seed  (4) 

Rice  flour 

Rice,  seed  (5) 

Rice  embryo  (5) 

Barley  bran  \Hordeum  xmlgarc)   (3) 

Barles",  grain  (4) 

Barley,  grain  (6)   (5) 

Barley  bran  (6)  (5) 

Barley  grain  minus  bran  (C)  (5) 

Barley,  grain  (Hordeum  distich.)  (2) 

Barley,  grain  (Hordeum  distich.)  (2) 

R3-e  flour 

Fye,  grain  (Sicale  ccreale)  (2) ... 

Millet  (Panicumfrumcntaceum)  (3) 

Sesame  seeds  (SesamUm  indicum)  (3) 

Castor  bean,  seed  (Ricinus  communis)  (3) 
Rape  seed  (Brassicanapus  oleifera)  (2)  ... 
Oil  cake  of  rape  (Brassica  napus)  (3)  . . 

Rape  seed  (Brassica  napus)  (4) 

Radish  root  juice  (7) 

Turnip  root  juice  (7) 

Apple,  juice  (7) 

Pear,  juice  (7) 

Corn,  grain  (4)  (5) 

Corn,  bran  (4)  (5) 

Corn,  germ   (4)  (5) 

Com,  endosperm  (4)  (5) 

Corn,  grain  (Zea  mays)  (2) 

Oat,  grain  (4)  (5) 

Oat,  hull  (4)   (5) . . . 

Oat,  grain  minus  hull  (4)  (5) 


Schulze,  Winterstein 

Posternak 

Vorbrodt 

Vorbrodt 

Posternak 

Vorbrodt 

Posternak 

Posternak 

Rising 

Posternak 

Rising 

Posternak 

Vorbrodt 

Vorbrodt 
Patten,  Hart 
Suzuki,  Yoshimura 
Suzuki,  Yoshimura,  Takaishi 

Vorbrodt 

Rising 
Suzuki,  Yoshimura 
Suzuki,  Yoshimura,  Takaishi 

Rising 

Bernardini 

Bernardini 
Suzuki,  Yoshimura 
Suzuki,  Yoshimura,  Takaishi 
Hart,  Tottingham 
Hart,  Tottingham 
Hart,  Tottingham 

Vorbrodt 

Vorbrodt 

Rising 

Vorbrodt  , 

Suzuki,  Yoshimura 
Suzuki,  Yoshimura 
Suzuki,  Yoshimura 

Vtrbrodt 
Suzuki,  Yoshimura 
Suzuki,  Yoshimura,  Takaishi 
Suzuki,  Yoshimura 
Suzuki,  Yoshimura 
Suzuki,  Yoshimura 
Suzuki,  Yoshimura 
Hart,  Tottingham 
Hart,  Tottingham 
Hart,  Tottingham 
Hart,  Tottingham 

Vorbrodt 
Hart,  Tottingham 
Hart,  Tottingham 
Hart,  Tottingham 


0.33 
0.60 
0.15 
0.07 
1.33 
0.11 
0.72 
0.26 
0.11 
0.42 
0.29 
0.25 
0.03 
0.02 
0.92 
0.58 

6.' 136 

0.08 
1.68 

o.'ii 

0.436 

5.14 

0.24 

6.'i9 
0.15 
0.17 
0.17 
0.12 
0.09 
0.12 
0.34 
0.13 
0.11 
0.20 
0.53 


0.13 
0.00 
0.13 
0.15 
0.17 
0.18 
0.09 
0.22 


91.46 
21.65 
14.39 
91.44 
15.00 
86.26 
70.80 
19.00 
81.60 
52.00 
82.60 
9.29 
4.39 
68.10 
52.00 
55.50 
29.90 
29.00 
74.17 
41.64 
69.00 
45.90 
82.90 
44.00 
56.55 
38.00 
68.20 
29.80 
36.40 
34.30 
25.00 
28.90 
44.97 
16.24 
41.61 
38.05 
44.46 
62.11 
15.13 
15.06 
48.14 
46.15 
44.80 
0.00 
34.20 
35.70 
48.90 
43.90 
21.90 
53.70 


piler. 


(1) 
(2) 


Computed.     Percent  of  fat-free  seeds.        NaCl  extract,  precipitated  hot. 

Organic  phosphorus  soluble  in  1.0  percent  acetic  acid;   computed  from  P20B  by  com- 


(3)  Percent  of  dry  substance  obtained  by  absolute  alcohol  precipitation  from   0.2   per- 
cent HC1  extract. 

(4)  Organic  phosphorus   soluble   in   0.2   percent  HC1. 

(5)  Percent  of  total  computed  by  compiler. 

(6)  Phosphorus  soluble   in   0.2  percent  HC1    (considering  that  the   inorganic  fraction   of 
this  is  small  enough  to  be  ignored). 

(7)  Total  organic  phosphorus  of  expressed  juice;   grams  per   100   c.c. 


PHOSPHORUS  METABOLISM  53 

To  quote  finely  from  Posternak  (1903a) : 

Phytin  is  stored  as  reserve  material  in  all  grains,  tubers,  rhi- 
zomes and  bulbs,  where  it  is  destined  for  the  development  of  the  em- 
bryo. Other  bodies  from  which  it  has  been  extracted  besides  those 
reported  in  this  paper  are — rape,  lupine,  wheat,  corn,  potatoes,  dahl- 
ia bulbs,  carrots  and  even  onions.  In  the  grains,  where  there  is 
very  little  mineral  phosphate,  it  forms  at  least  70-90  percent  of 
the  total  phosphorus. 

Anderson  (1912b)  was  unable  to  find  in  wheat  bran  any  of  the 
characteristic  salts  of  phytic  acid. 

The  purified  barium  salts  of  the  compounds  obtained  corre- 
sponded to  the  formulae:  C25H55054P9Ba5  and  C20H45O49P9Ba5.  The 
only  acid  that  could  be  isolated  was  C20H35O49P9,  and  all  of  the 
barium  salts  obtained  could  be  changed  into  salts  of  this  acid 
by  processes  which  liberate  reducing  substances.  Hence  it  was 
concluded  that  this  acid  is  the  only  organic-phosphoric  acid  present, 
and  that  wheat  bran  does  not  contain  phytin. 

Rather  (1912,  1913a,  1913b)  has  investigated  a  corresponding 
compound  obtained  from  cottonseed  meal.  It  was  his  conclusion 
that  this  and  the  wheat  bran  compound  are  identical,  and  that  the 
free  acid  from  both  is  represented  by  the  formula  C12H41P9Q42.  An- 
derson (1914a),  however,  thinks  that  Rather's  silver  salt  prepara- 
tions are  not  chemically  pure,  and  from  his  own  investigations 
(1912c,- 1914a)  believes  that  the  organic  phosphoric  acid  in  cotton- 
seed meal  must  be  inosite  hexaphosphoric  acid,  C6H18024P6,  or  some 
multiple  of  the  same.  Anderson  gives  the  same  formula  for  com- 
pounds obtainable  from  oats  (1914b),  from  corn  (1914c),  and,  final- 
ly, from  commercial  phytin  (1914d). 

Salts  of  Phytic  Acid.  Phytic  acid  forms  neutral  salts,  acid 
salts,  double  salts  and  acid  double  salts  of  the  alkalis,  alkaline  earths 
and  heavy  metals.  The  solubility  of  these  salts  decreases  in  the 
order  above  named.  The  magnesium  compounds  are  more  soluble 
than  the  calcium,  and  the  calcium  more  soluble  than  the  barium 
or  strontium.  They  are  all  more  soluble  in  cold  than  in  hot  water, 
and  heating  often  precipitates  them.  Several  such  salts  have  been 
isolated  from  plant  bodies;  and  Anderson  (1911)  has  added  largely 
to  the  number  by  artificial  production.  Elementary  analysis  of 
these  salts  throws  light  on  the  basicity  of  the  acid  and  the  relative 
number  of  C  and  P  atoms  in  the  molecule.  Rose  gives  us  a  table 
compiled  from  the  various  elementary  analyses  reported  in  the  lit- 
erature. Rose's  conclusion  is  that  the  relative  number  of  atoms  in 
the  molecule' corresponds  with  the  ratio  6C:6P;  and — "It  seems 


54  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

probable  that  the  molecular  weight  when  accurately  determined  will 
be  reported  as  714  or  will  differ  from  this  by  the  molecular  weight 
of  three  molecules  of  water.  The  molecule  seems  to  contain  twelve 
hydrogen  atoms  readily  separated  in  ionization,  six  of  which  are  ex- 
ceedingly reactive ;  the  remaining  hydrogen  atoms  gradually  dimin- 
ish in  activity  by  twos,  the  last  four  being  slow  to  enter  into  an  ex- 
change with  bases.  The  more  readily  formed  salts  are  therefore 
those  corresponding  to  an  octavalent  acid  and  the  other  common 
ones  are  in  six  and  tenvalent  combinations." 

Synthesis  of  Phytic  Acid.  Anderson  speaks  of  a  paper  by  Con- 
tardi,  which  we  have  not  seen  (Atti  R.  Accad.  dei  Lincei,  Roma  [5] 
19  1  23),  in  which  the  author  reports  having  prepared  the  hexa- 
phosphoric  acid  ester  of  inosite  by  heating  inosite  in  an  excess  of 
phosphoric  acid  in  a  stream  of  carbon  dioxide  at  160-1 65 °C.  The 
free  ester  obtained  from  this  preparation  was  said  to  be  identical 
with  phytic  acid.  Carre,  however,  (Bull.  Soc.  chim.  de  France  [4] 
9,  195)  repeating  these  experiments,  found  that  the  products  de- 
scribed by  Contardi  were  merely  mixtures  of  free  phosphoric  acid 
with  the  alcohol,  together  with  their  decomposition  products  mixed 
with  monobarium  phosphate.  Anderson  has  made  repeated  ef- 
forts to  synthesize  phytic  acid,  without  accomplishing  it;  nor  was 
he  able  to  synthesize  a  hexa-phosphoric  acid  ester  of  inosite.  A 
tetra-orthophosphoric  acid  ester,  a  di-pyrophosphoric  acid  ester  of 
inosite  and  a  di-inosite  tri-pyrophosphoric  acid  ester  were  obtained 
in  pure  form,  and  analyzed.  (Anderson,  1912a.)  Of  them  Anderson 
says:  "These  compounds  are  in  physical  and  chemical  properties 
very  similar  to  phytic  acid.  They  form  analogous  acid  salts  Which  in 
appearance  and  solubility  seem  almost  identical  with  salts  of  phytic 
acid.  Whether  esters,  such  as  above,  are  found  in  nature  is  at 
present  unknown.  It  is,  however,  not  impossible  that  a  part  of 
the  organically  bound  phosphorus  existing  in  plants  may  be  present 
in  such,  or  similar  forms." 

Constitution.  Winterstein  (1897,  1908),  repeatedly  obtaining 
a  cleavage  of  the  compound  into  inosite  and  phosphoric  acid,  be- 
lieved it  to  be  a  conjugated  inosite-phosphoric  acid.  Posternak 
(1903a,  1903d),  however,  was  of  the  opinion  that,  although  inosite 
is  obtained  in  such  cleavage,  the  grouping  present  in  the  phytin  is 
that  of  a  simpler  compound,  that  of  the  alcoholic  isomer  of  formal- 
dehyde, CH'OH,  and  that  when  this  is  set  free  from  the  acid  poly- 
merization occurs,  forming  the  inosite  by  union  of  six  such  groups. 
He  interpreted  phytin  as  being  the  first  product  in  the  organization 


PHOSPHORUS  METABOLISM  55 

of  inorganic  phosphorus  taking  place  under  the  influence  of  chlor- 
ophyll in  direct  sunlight.  He  called  it  anhydro-oxymethylene-di- 
phosphoric  acid  and  proposed  the  empirical  formula,  C2HsP209,  and 
the  structure  indicated  by  the  following : 

H 

/ 

CH— OPO(OH)2 

/ 
O 
\ 
CH— 0-PO(OH)2 
\ 
H 

The  analyses  of  Patten  and  Hart  (1904)  were  said  to  support 
this  formula. 

Suzuki,  Yoshimura  and  Takaishi  (1907),  because  of  enzymatic 
splitting  off  of  inosite,  decided  that  inosite  was  originally  present  in 
the  phytin,  and  that  the  substance  must  be  inosite  hexaphosphoric 
acid.  They  constructed  the  following  formula  to  represent  their 
view. 

HO  OH 

O— P— O— HC— CH— 0— P— 0 

/  \ 

HO         |   |        OH 

HO         ||        OH 

\  / 

O— P— 0— HC— CH— 0— P— 0 

HO^        ||       XOH 

HO  OH 

V  / 

0— P— 0— HC— CH— O— P— O 

. .  /  \ 

HO  OH 

The  discovery  by  Neuberg  (1908)  that  both  inosite  and  phytin 
yield  furfurol  when  distilled  with  phosphorus  pentoxide  and  phos- 
phoric acid,  respectively,  led  him  to  believe  that  the  inosite  ring  ex- 
ists already  formed  in  the  phytin,  and  he  suggested  the  following 
structural  formula : 


56  OHIO  EXPEEIMENT  STATION:  TECHNICAL  BUL.  § 

HO  OH 

HO\  H     H  /OH 

HO-^-P— O— C  —  C— 0— P— OH 

/  \ 

0  ||  0 

\  / 

HO— P— O— CH  HC— O— P— OH 

HO/  |        |  \^0H 

HO  HC  — CH  OH 

I        I 

o    o 

HO       I        |      OH 

\  / 

HO—  P      P  —OH 

/  \/\ 
HO         0        OH 

Levene,  (1909a)  (to  quote  from  Rose)  "working-  with  a  prepar- 
ation from  hempseed,  was  led  to  believe  that  the  'phytin'  of  this 
gram  contained  in  its  molecule  phosphate,  inosite  and  a  carbohy- 
drate of  the  pentose  group.  His  work  was  criticized  by  Neuberg 
(1909),  who  claimed  that  there  were  impurities  in  the  preparation. 
In  view  of  the  known  intimate  association  of  the  phytin  with  pro- 
tein and  carbohydrate  in  the  aleurone  grain,  and  the  possible  occur- 
rence of  a  chemical  combination  of  both  phyto-phosphate  and  carbo- 
hydrate with  protein,  it  is  conceivable  that  Levene  had  a  product 
holding  pentose  as  an  integral  part  and  not  as  an  impurity,  though 
in  view  of  all  the  available  evidence  Neuberg's  criticism  seems  at  the 
present  time  somewhat  justifiable." 

Starkenstein  (1910),  who  thinks  that  inosite  bears  some  defi- 
nite relation  to  the  phenomena  of  growth  of  animals  as  well  as  of 
plants,  believes  that  the  acid  to  which  phytin  corresponds  is  not  a 
simple  ester  but  a  complex  pyro-phosphoric  acid  compound,  and  that 
the  acid  salts  usually  resulting  from  reaction  with  divalent  metals 
are  to  be  explained  by  the  union  of  the  divalent  atoms  with  hydrox- 
yls  of  adjacent  phosphoric-acid  residues.  His  interpretation  would 
give  the  ratios  of  C:P:OH  as  6:6:12,  which  is  supported  by  most 
of  the  analyses.  Titration  with  uranium  acetate  shows  only  one 
half  of  the  true  phosphoric  acid  value,  because  only  one  half  of  the 
free  hydrogens  are  readily  reactive. 

The  analyses  of  Levene,  Vorbrodt  (1910)  and  Rising  (1910)  do 
not  support  any  of  these  formulae  as  to  the  relative  amounts  of  car- 
bon and  phosphorus ;  and  we  understand  that  those  of  Plimmer  and 
Page  do  not  (Chem.  Abs.). 


PHOSPHORUS  METABOLISM  57 

Anderson  (1914d)  has  succeeded  in  obtaining  from  commercial 
phytin  a  crystallizable  barium  salt,  and  the  acid  from  the  same, 
analysis  of  which  corresponds  with  the  formula  C2H608P2  or  C6H18 
024P6.  No  direct  evidence  has  been  gained  as  to  the  molecular  mag- 
nitude, but  the  second  formula  is  thought  to  be  the  more  probable. 
This  differs  from  Neuberg's  formula  for  phytic  acid  by  three  mole- 
cules of  water,  and  is  identical  with  inosite  hexaphosphate.  Ander- 
son, therefore,  believes  the  phytic  acid  to  be  either  inosite  hexa- 
phosphate or  an  isomer  of  the  same.  He  finds,  as  did  Starkenstein, 
that  only  one  half  of  the  acid  hydroxyls  are  particularly  reactive. 
Both  the  salts  and  the  free  acid  when  kept  at  ordinary  temperatures 
decompose  slowly,  with  liberation  of  inorganic  phosphate,  the  acid 
decomposing  much  faster  than  the  salt.  Inosite  was  not  found  among 
the  products  of  this  spontaneous  decomposition. 

Possible  relations  between  phytin  and  lecithin  (phosphatids) 
on  the  one  hand  and  nucleic  acids  on  the  other  have  been  pointed 
outbylljin  (1906),  Parrozzani  (1909)  and  Rising  (1910). 

Phytin-Splitting  Enzyme.  Suzuki,  Yoshimura  and  Takaishi 
(1907)  isolated  an  enzyme  from  rice  and  wheat  bran  which  splits 
phytin  into  phosphoric  acid  and  inosite,  and  which  is  probably  wide- 
spread in  the  plant  world.      They  named  the  new  enzyme  "phytase." 

A  part  of  Vorbrodt's  (1910)  study  was  with  regard  to  enzy- 
matic decomposition  of  the  organic  phosphorus  compounds  of  bar- 
ley and  corn.      He  concluded  that : 

"1.  The  organic  phosphorus  compounds  of  barley  and  corn 
grains,  both  those  soluble  in  1%  acetic  acid  and  those  insoluble,  can 
be  decomposed  under  the  influence  of  enzymes,  in  which  process 
mineral  phosphoric  acid  is  split  off. 

"3.  Temperature  exerts  a  decided  influence  on  the  progress  of 
the  decomposition  of  the  organic  phosphorus  compounds;  a  little 
above.  0°  there  is  no  decomposition  of  the  soluble  organic  phosphorus 
compounds  of  barley,  the  optimum  is  about  28°C,  above  48°  the 
progress  of  decomposition  is  very  slow. 

"4.  The  enzymes  which  split  the  organic  phosphorus  com- 
pounds are  already  present  in  the  barley  grain,  but  in  the  corn 
grain  the  amount  is  very  insignificant ;  they  develop  in  large  amount 
during  germination,  when  they  are  present  more  abundantly  in  the 
germ  and  the  scutellum  than  in  the  rest  of  the  seed. 

McCollum  and  Hart  (1908)  found  evidence  of  such  a  phytin- 
splitting  enzyme  in  the  liver  and  the  blood  of  the  calf,  but  not  in  the 
muscle  or  kidney. 

Jegorow  (1912)  doubts  the  existence  of  phytase,  since  under 
the  conditions  of  the  work  by  Suzuki  phosphoric  acid  splits  off  with- 
out the  enzyme  preparation.  He  finds  phytin  quite  unstable.  Ino- 
site and  phosphoric  acid  result  from  boiling  with  water. 


58  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

PHOSPHATIDS 

INTBODUCTION 

Our  present  knowledge  of  the  phosphatids  has  been  attained 
largely  through  the  systematic  study  of  the  composition  of  the 
brain.  Of  these  studies  the  most  significant  as  supplying  a  basis 
for  the  classification  of  phosphatids  (originating  the  name  "phos- 
phatid")  was  that  of  Ludwig  W.  Thudichum  (1901).  Thudichum's 
earlier  work  (1875)  and  that  of  Gutnikov  (1896-7)  should  also  be 
mentioned.  Other  more  recent  analyses  and  general  discussions 
of  the  brain  which  include  classifications  of  the  phosphatids  are 
those  of  Waldemar  Koch  (1904),  of  Coriat  (1905)  and  of  Frankel 
(1909a).  The  basis  of  classification  which  Thudichum  introduced 
has  proved  to  be  very  satisfactory  and  has  continued  in  use  with 
but  slight  modifications. 

Since  the  phosphatids  resemble  the  fats  in  their 
solubilities,  and  in  some  other  properties,  it  has  been  found 
convenient  to  discuss  these  groups  together  with  cholesterins  and 
cerebrosides  under  the  comprehensive  name  of  "lipoids,"  or  "fat- 
like substances."  Ivar  Bang  defines  lipoids  as,  "compounds  which 
are  soluble  in  organic  solvents  such  as  ether,  alcohol,  chloroform  and 
benzol." 

Bang's  book,  published  in  1911,  is  freely  used  in  the  preparation 
of  this  article  and  his  classification  is  followed.  Rosenheim  (1909) 
presented  certain  "Proposals  for  the  Nomenclature  of  the  Lipoids," 
in  which  several  names  are  discarded,  and  the  group  divisions  are 
fitted  to  those  which  are  retained.  The  classification  does  not  ma- 
terially differ  from  that  of  Bang. 

The  phosphatids  are  in  every  way  the  most  important  lip- 
oids. They  are  of  great  physiological  interest,  both  as  food  con- 
stituents of  specific  importance  and  as  cell  constituents  with  signif- 
icant functions.  Apparently  they  are  primary  constituents  of 
cells,  that  is,  are  found  in  all  cells,  and  are  necessary  for  the  life  of 
the  cell.  They  are  interesting  in  their  chemical  relations  and  are 
as  difficult  to  obtain  pure  as  are  the  proteins. 

The  name  "phosphatid"  was  proposed  by  Thudichum  to  signify 
that  we  have  here  bodies  similar  to  phosphates  though  not  just 
like  them.  Phosphoric  acid  was  considered  to  be  the  central  group 
of  the  molecule,  with  which  are  combined  an  alcohol,  fatty  acids  and 
a  nitrogenous  base.  The  alcohol  is  at  least  nearly  always  glycerin, 
and  the  base  is  mainly  choline.  If  there  be  one  such  phosphoric 
acid  molecule  only,  the  compound  is  a  monophosphatid ;  two  such, 
united,  may  form  a  diphosphatid.      Further,  there  may  be  one  or 


PHOSPHORUS  METABOLISM  59 

two,  perhaps  three,  possibly  more  nitrogenous  groups  making  mon- 
amino,  diamino,  etc.,  compounds.  The  phosphatids  are  classified 
on  this  basis.  Thus,  if  on  elementary  analysis  a  substance  of  this 
class  is  shown  to  have  the  relative  number  of  atoms  of  nitrogen  and 
phosphorus  such  that  N:P=2:1,  it  is  placed  in  the  diamino-mono- 
phosphatid  group.  These  names  are  still  used  although  the  pres- 
ent conception  makes  glycerin  more  centrally  significant  in  the  mole- 
cule than  Thudichum  thought  it  to  be. 

Bang's  classification  makes  use  of  a  difference  pointed  out  by 
Frankel  (1909b)  by  which  the  phosphatids  which  contain  an  un- 
saturated fatty  acid  form  one  class,  and  those  which  contain  only 
saturated  fatty  acid  radicals  form  another.  The  former,  in  distinc- 
tion from  the  latter,  are  semisolid  or  viscous ;  they  do  not  crystal- 
lize; they  oxidize  in  the  air,  they  are  easily  decomposed  and  they 
readily  react  with  other  substances.  They  are  for  these  reasons 
particularly  difficult  to  obtain  pure,  and  doubtless  most  of  the  prep- 
arations that  have  been  examined  have  been  mixtures.  Accord- 
ing to  Erlandsen,  the  cadmium  chloride  method  of  preparation 
causes  some  decomposition. 

The  following  is  Bang's  classification  including  the  best-known 
phosphatids : 

BANG'S  CLASSIFICATION  OF  PHOSPHATIDS 

A.  Unsaturated  Phosphatids 

I.  Monamino-monophosphatids,  (N:P,  1:1) 
Lecithin,  cephalin  and  myelin. 

II.  Monamino-diphosphatids,   (N:P,  1:2) 

Cuorin,  liver  phosphatid  and  monamino-diphosphatid 
from  egg-yolk. 

III.  Triamino-diphosphatids,  (N:P,  3:2) 

B.  Saturated  Phosphatids 

.   I.     Diamino-monophosphatids,  (N:P,  2:1) 

Sphingomyelin  and  diamino-phosphatids  from  muscle, 
from  egg-yolk  and  from  horse  pancreas. 

II.  Triamino-monophosphatids,  (N:P,  3:1) 
Neottin  and  carnaubon. 

III.  Protagon. 

C.  Insufficiently  Characterized  Phosphatids 

D.  Plant  Phosphatids 


60  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

A.    UNSATURATED  PHOSPHATIDS 

1.     MONAMINO-MONOPHOSPHATIDS 
LECITHIN,  OR  LECITHINS 

Occurrence.  The  best  known  of  the  phosphatids  and  the  one 
which  is  apparently  most  widely  distributed  in  the  animal  kingdom 
is  lecithin.  However,  it  is  now  recognized  that  not  all  of  the  sub- 
stances which  have  been  reported  as  lecithin  should  be  given  this 
name.  It  is  only  within  quite  recent  years  that  sufficiently  dis- 
criminating work  has  been  done  to  differentiate  lecithin  from  cer- 
tain other  phosphatids.  It  is  possible  that  it  is  a  primary  con- 
stituent of  animal  cells,  but  it  is  not  impossible  that  it  is  sometimes 
lacking  and  its  place  taken  by  another  phosphatid.  Apparently  it 
is  not  present  in  any  vegetable  substance.  Its  identity  is  little  to 
be  questioned,  however,  as  found  in  heart  muscle  (Erlandsen,  1906, 
1907 ;  MacLean,  1908a) ,  muscle  of  the  extremities  (Erlandsen,  1906, 
1907),  egg-yolk  (Stern  and  Thierf elder,  1907 ;  Erlandsen,  1907;  Mac- 
Lean,  1909c)  and  liver  (Baskoff,  1908). 

MacLean  (1912b),  using  special  purification  processes,  proves 
the  principal  phosphatid  of  kidneys  and  of  muscle  to  be  at  least  lec- 
ithin-like in  its  nitrogen  and  phosphorus  content. 

The  discovery  of  lecithin  was  made  by  Gobley,  who  found  it  in 
egg-yolk  in  1846,  in  the  brain  of  hen,  sheep  and  man  in  1847,  in  the 
eggs  and  milt  of  carp  in  1850,  in  the  blood  of  ox  and  man  in  1851, 
and  in  bile  in  1856.  He  more  fully  studied  that  in  human  brain  in 
1877.       (Gobley,  1846,  1847,  1850a,  1850b,  1851,  1856,  1877.) 

Constitution.  Gobley  recognized  the  existence  in  lecithin  of 
glycerophosphoric  acid,  of  fatty  acid,  and  of  nitrogen  in  an  NH3 
group.  The  nitrogenous  group  was  thought  by  Liebreich  (1865)  to 
be  neurine,  but  Strecker  (1868)  showed  it  to  contain  oxygen  and  to 
be  choline.      It  is  now  recognized  as  being  mainly  choline. 

C8H  —  OH 

N=(CH3)3 

\ 

OH 

(See  also  Bergell,  1900,  1901.) 

MacLean  (1908a,  1909a,  1909c)  has  shown  that  not  all  of  the 
nitrogen  in  lecithin  is  present  as  choline,  and,  moreover,  that  leci- 
thins from  different  sources  do  not- agree  in  the  proportion  of  choline 
they  contain.      Very  careful  examination  of  lecithin  from  heart 


PHOSPHORUS  METABOLISM  61 

muscle  showed  not  more  than  42.6  percent  Of  the  nitrogen  to  be 
present  as  choline,  while  in  commercial  lecithin  80  percent  and  in 
that  from  egg-yolk  66  percent  of  the  nitrogen  was  present  in  this 
form.  MacLean  was  led  to  feel  that  some  of  the  nitrogen  must  be 
in  the  form  of  amino  acid.  Trier  (1912,  1913b)  has  isolated 
from  the  hydrolytic  products  of  egg  lecithin,  as  well  as  from  related 
compounds  of  plant  origin,  a  compound  of  amino-ethyl-alcohol  which 
he  thinks  may  be  the  mother  substance  of  choline,  and  which  he 
names  "colamin." 

The  structure  of  the  lecithin  molecule  was  early  studied  by 
Diaconow  (1867b,  1868a,  1868b),  by  Strecker  (1868)  and  by  Gilson 
(1888).  Diaconow  held  that  the  choline  is  bound  with  the  phos- 
phoric acid  in  salt-like  union,  while  Strecker  and  Gilson  believed  that 
it  is  ester-like.  The  constitution  suggested  by  Strecker  is  practi- 
cally that  which  is  now  generally  accepted. 

OH 

CH2— O— P— O— C2H4 

I  II  \     . 

CHOR      O    (CH3)3=N 

I  / 

CH8OR  OH 

In  this  formula  R  stands  for  a  fatty  acid  radical.  Two  of 
these  radicals  are  supposed  to  take  the  place  of  the  hydrogen  in  two 
of  the  OH  radicals  of  the  tribasic  alcohol,  glycerin,  and  the  phos- 
phoric acid  that  of  the  other.  This  formula  is  supported  by  the 
observations  on  optical  activity  as  made  by  Ulpiani  (1901)  and  by 
Willstatter  and  Ludecke  (1904),  but  not  by  the  recent  study  of  Mal- 
engreau  and  Prigent  (1912)  on  the  rate  of  cleavage  of  the  different 
radicals  in  acid  hydrolysis.  According  to  these  observations,  the 
splitting  off  of  choline  and  of  the  fatty  acids  takes  place  at  the 
same  time,  and  much  more  rapidly  than  the  phosphoric  acid  is  freed 
from  its  glycerophosphoric  acid  combination.  Hence  it  is  thought 
that  the  linkings  of  the  phosphoric  acid  with  other  residues  (gly- 
ceryl and  choline)  are  not  of  the  same  nature,  as  they  would  be 
according  to  the  above  formula. 

Of  the  phosphatid  components  the  fatty  acids  are  of  most  val- 
ue in  distinguishing  one  phosphatid  from  another.  Thudichum 
thought  that  oleic  acid  is  characteristic  of  lecithins,  and  that  it  is 
associated  with  some  other,  usually  palmitic  or  margaric  acid.  Hen- 
riques  and  Hansen  (1903)  showed  that  there  must  be  some  acid 
present  more  unsaturated  than  is  oleic  acid.  Cousin  (1903)  and 
Erlandsen  (1906,  1907)  both  speak  of  linoleic  acid  as  a  phosphatid 


62 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


constituent.  Attempts  to  identify  the  acid  by  its  iodine  number 
have  brought  results  which  cannot  at  present  be  reconciled.  Of 
course  oxidation,  which  so  easily  takes  place,  rapidly  reduces  the 
iodine  number.  Bang's  conclusion,  after  considering  all  the  evi- 
dence, is  that  there  are  in  lecithin  two  fatty  acids,  one  of  which  is 
stearic  and  the  other  an  as  yet  undetermined  unsaturated  acid. 
The  saturated  acid  may  be  palmitic  (Serono  and  Palozzi,  1911). 

These  disagreements  and  uncertainties  with  regard  to  the  con- 
stituents of  lecithin  are  doubtless  due  in  part  to  the  fact  that  the 
pure  compound  has  not  been  obtained,  since  lecithin  so  readily  oxi- 
dizes, and  is  so  difficult  to  free  from  other  phosphatids,  fats,  etc. 
It  is  quite  doubtful  if  the  substances  examined  were  individual  com- 
pounds. Bang  says  that  elementary  analysis  has  shown  Merck's 
preparation  to  be  quite  impure  and  unsuited  to  scientific  work ;  and 
that  the  preparation  of  the  proprietary  product  known  as  "agfa-leci- 
thin"  by  Bergell's  method,  which  makes  use  of  the  CdCl2  precipita- 
tion, has  been  shown  by  Erlandsen  to  cause  a  serious  decomposition. 
Hence  conclusions  which  have  been  drawn  on  the  basis  of  work  with 
these  products  are  of  little  value.1  Bang  considers  that  no  suitable 
method  of  preparation  has  been  described  except  that  of  Erlandsen. 

It  cannot  be  said,  finally,  whether  there  is  but  one  lecithin  or 
whether  there  are  several  lecithins  isolated  from  animal  products  by 
the  same  process. 

Elementary  Analysis.  The  elementary  analyses  and  empirical 
formulae  reported  by  different  workers  are  nearly  alike,  although 
not  identical.      A  few  are  quoted  here : 

ELEMENTARY  ANALYSES  OF  LECITHINS— Percent 


Author 


Thudichum 

Koch 

Erlandsen 

MacLean 

Erlandsen 

Stern  and  Thierfelder 

Mac  Lean 

Baskoff 


Date, 
refer- 
ence 

Source  of 
lecithin 

C 

II 

N 

P 
4.00 

» 

Brain 

66.75 

18.67 

1.81 

1902b 

Brain 

64.03 

10.4 

1.8 

3.79 

1906, 

1907 
1908a 

Muscle  of 
heart 

Muscle  of 
heart 

66.29 
66.27 

10.17 
10.32 

1.87 
1.85 

3.95 
3.975 

1907 

Muscle  of 
thigh 

65.96 

10.20 

1.82 

3.93 

1907 

Egg-yolk 

64.63 

10.96 

2.08 

3.97 

1909a 

Egg-yolk 

64.18 

10.6 

1.876 

3.95 

1908 

Liver 

64.64 

10.71 

1.95 

4.00 

Formula 


C13H&1NP08(1) 


C43H8oNP09 
CsHsoNPOs 


(J)      Quoted  through  Bang   (1911). 

(1)  Altchul's  (1912)  assertion  that  "Agfa-lecithin"  is  not  made  by  Bergell's  method, 
but  by  a  method  worked  out  by  himself,  which  gives  a  pure  product,  is  answered  by  Bang 
(1912)   by  saying  that  he  based  his  statement  as  to  the  method  used  on  the  authority  of  the 

"Agfa"    company. 


PHOSPHORUS  METABOLISM  63 

Properties.  Lecithin  is  a  white,  yellow,  or  orange-colored  sub- 
stance, usually  amorphous,  and  of  a  waxy  consistency  when  dry. 
Some  have  obtained  it  in  crystalline  form,  the  crystals  being  thin 
flakes.  It  is  very  hygroscopic,  and  in  the  air  it  takes  up  moisture,  and 
softens  to  a  semifluid  mass.  It  also  readily  takes  up  oxygen  from 
the  air,  and  in  doing  so  grows  darker  in  color.  Unlike  the  fats,  lec- 
ithin becomes  wet  and  sinks  in  water.  Gradually  it  takes  up  water 
and  swells  to  a  gelatinous  mass,  and  finally  spreads  to  an  opaque, 
colloidal  solution.  Lecithin  is  precipitated  from  this  colloidal  so- 
lution by  certain  acids  and  salts,  and  precipitation  is  prevented  by 
other  salts.  It  is  thought  that  the  tendency  of  phosphatids  in  gen- 
eral to  pass  into  the  colloidal  state,  and  their  properties  as  colloids, 
may  be  significant  in  their  functioning  within  the  cells.  The  pre- 
cipitating power  seems  to  be  due  to  the  individual  ions  and  depend- 
ent on  their  concentration.  It  is  a  reversible  reaction,  and  the  total 
effect  is  a  summation  of  the  effects  of  the  individual  ions  present. 
Quantitative  studies  of  such  precipitations  of  lecithin  and  other 
lipoids  have  been  made  by  W.  Koch  (1902a,  1903,  1907a,  1909b, 
1910b),  Koch  and  MacLean  (1910),  Koch  and  Mostrom  (1910),  Koch 
and  Pike  (1910),  Koch  and  Williams  (1910),  Porges  and  Neubauer 
(1907),  Long  (1908),  Long  and  Gephart  (1908a)  and  Feinschmidt 
(1912).  See  also  the  physicochemical  studies  of  Handovsky  and 
Wagner  (1911)  concerning  the  reactions  of  such  emulsions  with  in- 
organic salts  and  with  proteins.   . 

Lecithin  is  easily  dissolved  by  alcohol,  either  hot  or  cold,  and 
is  precipitated  from  this  solution  by  water.  It  is  soluble  also  in 
ether,  chloroform,  carbon  bisulphide,  benzene  and  some  other  or- 
ganic solvents,  but  nearly  insoluble  in  cold  acetone  and  in  methyl 
acetate.  The  power  of  lecithin  (and  other  lipoids)  in  such  organic 
solvents,  to  take  up  organic  substances,  has  recently  been  investi- 
gated by  Loewe  (1912a,  1912b,  1912c,  1912d)  and  is  attributed  to 
adsorption.  R.  Cohn  (1913)  thinks  that  on  longer  action  the  ad- 
sorption becomes  so  intensive  that  probably  a  chemical  adsorption 
results. 

According  to  Ulpiani  (1901),  and  to  Willstatter  and  Liidecke 
(1904),  lecithin  shows  by  its  optical  activity  that  it  has  an  asym- 
metric molecule.  Paul  Mayer  (1905,1906b)  separated  the  racemic 
form  under  the  influence  of  an  enzyme  (lipase)  into  dextro-  and 
laevo-lecithin,  which  then  behaved  differently  toward  the  lipase,  the 
laevo-lecithin  remaining  unchanged  while  the  dextro-lecithin  broke 
down  into  fatty  acids  and  dextro-glycerophosphoric  acid. 


64  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Lecithin  is  readily  saponified  by  alkalis  and  especially  by 
baryta,  yielding  fatty  acids,  glycerophosphoric  acid  and  choline. 
The  cleavage  with  dilute  acids  is  quite  slow.  Casanova's  (1911) 
observations  indicate  that  the  first  step  in  the  cleavage  resulting 
from  the  taking  up  of  water  in  the  presence  of  oxygen  is  a  separa- 
tion of  choline  and  glycerophosphoric  acid  esters  of  the  fatty  acids, 
further  taking  up  of  water  being  required  for  the  cleavage  of  these 
esters.  Malengreau  and  Prigent  (1912)  found  that  only  about  70 
percent  of  the  choline  and  fatty  acids  was  split  off  in  two  hours  by 
0.1N  sulphuric  acid,  that  after  13%  hours  they  were  nearly  all  split 
off,  but  the  cleavage  of  the  glycerophosphoric  acid  was  much  more 
slow,  and  was  still  incomplete  after  72  hours.  Lecithin  is  cleaved 
by  digestive  ferments. 

Compounds.  Lecithin  shows  more  power  to  enter  into  com- 
bination with  other  substances  than  do  most  organic  compounds. 
These  combinations  are  of  different  kinds;  sometimes  additions  in 
molecular  relation,  forming  well-characterized  compounds,  and 
sometimes  in  other  combinations  of  which  the  nature  is  not  known, 
which  may  be  chemical  complexes  or  may  be  adsorption  products. 
Thudichum  suggests  that  it  may  be  of  considerable  physiological 
significance  that  the  phosphatids  generally  react  with  so  many  kinds 
of  substances  and  are  thereby  changed,  and  that  these  reactions' 
are  so  dependent  on  the  concentrations  of  the  reacting  substances. 

Combinations  of  lecithin  with  inorganic  acids,  with  some  bases, 
and  with  both  inorganic  and  organic  salts  have  been  studied.  These 
seem  to  be  usually  chemical  and  perhaps  mainly  addition  products, 
though  it  has  been  shown  in  some  cases  that  partial  decomposition 
accompanies  the  taking  up  of  salt. 

Bang  gives  the  following  summary  of  the  classes  of  organic 
compounds  with  which  lecithin  combinations  have  been  made;  (1) 
glucosides,  such  as  phlorizin,  salicin  and  amygdalin,  (2)  alkaloids, 
such  as  chlorides  of  morphine  and  nicotine  and  sulphate  of  strych- 
nine, (3)  toxins,  as  cobra  poison  and  bee  poison,  (4)  cholesterin,  (5) 
enzymes,  (6)  dyes,  (7)  carbohydrates,  (8)  proteins.  These  substan- 
ces have  not  been  very  well  studied  as  yet  and  only  the  last  two  need 
be  considered  here. 

Compounds  of  Lecithin  and  Carbohydrate.  Apparent  combina- 
tions of  lecithin  and  carbohydrate  have  been  prepared.  For  ex- 
ample, if  an  alcoholic  solution  of  lecithin  and  glucose  be  evaporated 
to  dryness,  a  substance  is  obtained  which  shows  different  solubilities 
from  either  component,  which  is  taken  as  indication  that  a  chemical 
union  has  taken  place.      Similar  substances  are  said  to  have  been 


PHOSPHORUS  METABOLISM  65 

isolated  from  organs  and  from  blood.  These  preparations  give  glu- 
cose on  decomposition,  also  the  cleavage  products  of  lecithin;  but 
as  they  are  not  constant  in  composition,  nor  permanent  in  their  solu- 
tions, or  under  repeated  evaporation,  it  seems  probable  that  no 
chemical  union  exists  but  that  we  have  here  simply  an  example  of 
the  power  lecithin  shows  to  change  by  its  presence  the  solubility  of 
other  compounds.  These  artificial  preparations  have  been  spoken 
of  as  identical  with  Drechsel's  jecorin,  but  that  idea  must  immedi- 
ately be  given  up  when  it  is  shown  that  jecorin  always  contains  sul- 
phur and  sodium.  However,  jecorin  may  be  also  a  similar  loose 
combination,  or  mixture  of  sugar  with  a  phosphatid  or  its  decompo- 
sition products. 

Jecorin  was  first  found  by  Drechsel  (1886),  and  has  usually 
been  obtained  by  the  use  of  his  method.  Drechsel  isolated  it  from 
the  liver  of  a  horse,  and  the  following  year  Baldi  (1887)  found  it  in 
the  liver  of  the  rabbit  and  the  dog,  in  beef  spleen,  horse  blood,  horse 
muscle  and  human  brain.  He  said  it  was  present  in  largest  amount  in 
liver  and  next  in  spleen.  Since  then  it  has  repeatedly  been  isolated 
from  liver  (P.  Manasse,  1895;  Meinertz,  1905;  Siegfried  and  Mark, 
1905 ;  P.Mayer,  1906a ;  Mark,  1907 ;  Waldvogel  and  Tintemann,  1906 ; 
Baskoff,  1908,  1909a,  1909b)  and  from  blood  (Henriques,  1897; 
Bing,  1899;  P.  Mayer,  1906a),  and  is  also  reported  from  adrenals 
(P.  Manasse,  1895),  spleen  (Waldvogel  and  Tintemann,  1906)  and 
bone  marrow  (Otolsky,  1906).  Both  smooth  and  striated  muscles 
showed  a  very  little  jecorin  in  Erlandsen's  (1906,1907)  analyses. 

The  nature  of  jecorin  is  not  established.  All  the  preparations 
examined  contained,  in  acTdition  to  phosphorus  and  other  constitu- 
ents of  lecithin,  sulphur  and  sodium  and  a  reducing  substance 
which  has  been  identified  as  glucose.  Bing  (1899)  looked  upon  it 
as  a  compound  of  lecithin  and  glucose,  and  it  has  been  investigated 
as  such.  However,  it  cannot  be  simply  such  a  cohipound,  on  ac- 
count of  the  sulphur  and  the  sodium  content,  as  has  been  said,  and 
it  is  doubtful  if  the  lecithin  is  chemically  united  with  the  glucose.  It 
may  be  that  the  union  of  the  reducing  substance  with  the  lecithin- 
like fraction  or  fractions  takes  place  at  the  time  of  their  simultan- 
eous precipitation.  At  least,  it  seems  probable  that  the  various 
substances  that  have  been  obtained  are  not  identical,  for  they  differ 
in  elementary  composition,  and  contradictory  properties  have  been 
reported. 

Baskoff  (1908),  after  a  review  of  the  subject,  and  careful  ex- 
perimental investigation  of  Drechsel's  jecorin  purified  in  several 
ways,  and  a  consideration  of  other  products  separated  during  the 


66  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

processes,  concludes  that  "Drechsel's  jecorin  may  be  characterized 
as  a  lecithin-like  complex  with  about  14  percent  glucose,  which  also 
always  contains  sulphur  and  an  inorganic  substance,"  and  that  there 
are  other  jecorin-like  bodies  not  identical  with  it.  The  close  sim- 
ilarity in  chemical  composition  of  his  several  purified  products 
argues  against  such  an  indefinite  mixture  of  substances  as  Paul 
Mayer,  Meinertz,  Siegfried  and  Mark,  and  A.  Mayer  and  Terroine 
have  suggested.  (References  as  previously  given  and  A.  Mayer 
and  Terroine,  1907.) 

Baskoff  finds  the  relation  of  N  :P=2 :1,  corresponding  to  a  diam- 
ino-monophosphatid.      Others  have  found  different  ratios. 

Lecithin-Protein  Compounds,  or  Lecitho-Proteins.  The  vitellin 
isolated  from  egg-yolk  contains  considerable  lecithin  (or  other  phos- 
phatid)  from  which  it  is  freed  with  great  difficulty,  and  there  is 
some  question  as  to  whether  it  should  not  be  looked  on  as  a  phos- 
phatid-globulin.  However,  there  seems  to  be  sufficient  evidence 
that  not  all  of  the  phosphorus  is  present  as  phosphatid  but  that  we 
have  here  a  nucleoalbumin,  still  containing  phosphorus  after  all 
phosphatid  is  removed.  Hoppe-Seyler  and  others  consider  that  the 
original  vitellin  is  chemically  combined  with  lecithin  and  that  the 
phosphatid-free  vitellin  is  therefore  to  be  looked  on  as  a  denatured 
compound.  According  to  T.  B.  Osborne  and  Campbell  (1900b),  the 
so-called  ovovitellin  is  a  mixture  of  several  vitellin-lecithin  combina- 
tions containing  15-30  percent  of  lecithin.  The  chemical  union  is 
not  proved,  nor  is  the  identity  of  the  lecithin  present,  which  may  be 
the  diamino-phosphatid  which  Erlandsen  found  in  egg-yolk. 

Other  lecithin-protein  compounds  have  been  discussed  under 
the  name  of  lecithalbumin.  Liebermann  (1891a,  1891b,  1893a, 
1893b)  isolated  what  he  called  lecithalbumins  from  the  mucous 
membrane  of  the  stomach  (hog),  from  kidney  (sheep),  liver  (lamb), 
lungs  and  spleen.  The  methods  used  in  preparing  and  handling 
these  substances  were  not  sufficiently  discriminating  to  prove  their 
nature.  Several  observers  think  that  the  phosphatids  of  the  serum 
and  of  chyle  are  in  combination  with  globulin. 

According  to  Erlandsen's  interpretation  of  his  extractions  from 
muscle,  the  lecithin  found  there  is  entirely  free  or  in  easily  cleavable 
condition,  while  the  diamino-phosphatids  are  in  some  complex  union 
with  proteins  or  other  substances  so  that  they  cannot  be  extracted 
until  the  proteins  have  been  precipitated  with  alcohol.  Micro- 
scopical observations  indicate  that  the  phosphatids  are  in  some  kind 
of  union  with  other  cell  constituents. 


PHOSPHORUS  METABOLISM  67 

Bing  (1901)  discusses  the  solubilities  of  a  variety  of  lecithin 
compounds.  For  an  extended  general  review  of  literature  on  leci- 
thin see  Merck  (1912). 

CEPHALIN 

Cephalin  was  first  studied  as  isolated  from  brain  by  Thudichum 
(Thudichum,  1875;  Thudichum  and  Kingzett,  1876),  and  has  since 
been  reported  by  W.  Koch  and  his  associates  as  found  in  various  tis- 
sues and  organs,  and  in  milk  and  yeast,  by  Stern  and  Thierfelder  in 
egg-yolk,  and  by  Cousin,  Falk  and  Frankel  and  associates  in  brain 
and  nerves.  (W.  Koch,  1902b,  1903,  1904,  1906;  W.  Koch  and 
Goodson,  1906;  W.  Koch  and  Woods,  1905;  W.  Koch  and  Mann, 
1907;  Stern  and  Thierfelder,  1907;  Cousin,  1906;  Falk,  1908; 
Frankel  and  Neubauer,  1909 ;  Frankel  and  Dimitz,  1909.)  Erland- 
sen  did  not  find  it  in  either  kind  of  muscle.  According  to  Thudi- 
chum, it  is  the  principal  phosphatid  in  the  brain,  and  Falk's  analysis 
of  peripheral  nerves  shows  considerably  more  cephalin  than  lecithin. 
It  is  doubtful  if  cephalin  has  ever  been  obtained  in  the  pure  state. 
Koch  reports  that  "agfa"  lecithin  is  two-thirds  cephalin.  Koch, 
however,  at  a  later  date,  abandoned  the  method  which  served  as  the 
basis  for  the  quantitative  separation  of  lecithin  and  cephalin. 
(Jour.  Am.  Chem.  Soc.  31, 1909,  p.  1349.) 

Cephalin  is  a  highly  unsaturated  monamino-monophosphatid.  It 
is  autooxidizable.  The  nitrogenous  base  is  probably  not  choline, 
but  a  base  of  lower  CH3  content  (W.  Koch,  1902b ;  Frankel  and  Neu- 
bauer, 1909). 

Baumann  (1913)  reports  that  this  base  exists  in  cephalin  as  a 
primary  amino  group,  and  Renall  (1913)  recognized  in 
cattle-brain  cephalin  an  amino  ethyl  alcohol,  which  he  con- 
siders a  characteristic  constituent  of  cephalin. 

The  base  is  united  with  glycerophosphoric  and  fatty  acids,  the 
latter,  according  to  Cousin  (1906)  containing  saturated  acids,  al- 
most entirely  stearic,  and  unsaturated  acids  of  the  linoleic  series. 
Cousin  worked  with  the  brain  of  cattle. 

Frankel  and  Dimitz  (1909)  think  that  the  cephalin  of  brain  is 
a  mixture  of  palmityl-cephalin  and  stearyl-cephalin,  with  more  of 
the  former.  Parnas  (1913)  states  that  all  of  the  fatty  acid  cleaved 
from  cephalin  by  baryta  is  stearic  acid.  Falk  (1908)  says  that  the 
cephalin  of  the  peripheral  nerves  is  not  the  same  as  that  of  the 
brain. 

Thudichum  gave  the  name  cephalic  acid  to  the  unsaturated 
acid,  which  he  considered  the  distinguishing  component  of  cephalins, 


68 


OHIO  EXPERIMENT  STATION!  TECHNICAL  BUL.  5 


and  which  through  its  marked  variability  in  composition  imparts 
variability  to  cephalins.  He  also  found  a  second  base.  Some  in- 
vestigators have  found  inorganic  base  elements,  as  Ca  or  K,  which 
seemed  to  belong  to  the  cephalins.  The  solubilities  of  the  prepar- 
ations reported  by  different  workers  do  not  agree. 

Some  of  the  elementary  analyses  reported  are  as  follows: 


ELEMENTARY  ANALYSES  OF  CEPHALIN— Percent 


Author 

Date    and 
Reference 

Source  of 
Cephalin 

C 

H 

N 

P 

Formula 

Koch    

1901 

1902b 
1909 
1908? 
1907 

Brain 
Brain 
Brain 
Nerve 
Egg-yolk 

60.00 

59.5 

62.05 

57.0 

59.68 

9.38 

9.8 

9.85 

9.1 

9.74 

1.68 
1.75 
1.69 
1.94 
1.57 

4.27 

3.83 

3.45 

4.4 

3.64 

C42H79NFOi3 
C42H82NPO13 

Frankel  and  Neubauer . 
Falk   * 

Stern    and    Thierfelder 

C1)      These  figures   are   taken  through  Bang    (1911a). 

Bang  gives  as  the  probable  formula,  C42H80NPO9 ;  Hammarsten 
gives  C42H82NP013.  If  there  are  several  cephalins,  the  formulas 
and  percentage  compositions  vary  accordingly. 

OTHER  MONOPHOSPHATIDS 

Other  monophosphatids  which  may  be  mentioned  here  are  the 
cephalin  variations,  the  paramyelin  (C38H75NP09)  and  the  myelins 
(C40H75NPO10)  of  Thudichum,  a  monophosphatid  found  by  Erland- 
sen in  the  acetone  solution  from  heart  muscle,  and  vesalthin  ob- 
tained by  Pari  (Frankel  and  Pari,  1909 ;  Frankel,  Linnert  and  Pari, 
1909)  as  the  CdCl2  salt,  C32H63NP09.CdCl2,  from  the  corresponding 
extract  from  pancreas. 

II     MONAMINO-DIPHOSPHATIDS 

According  to  Erlandsen,  diphosphatids  are  regularly  found  in 
large  amount  in  heart  muscle,  and  only  in  small  amount  in  the  stri- 
ated muscle.  One  or  two  have  been  found  elsewhere  in  the  animal 
body,  but  none  have  been  fully  studied.  They  have  not  been  found 
in  the  brain. 

CUORIN 

For  cuorin,  found  in  heart  muscle,  Erlandsen  gives  the  formula 
C71H125NP2021,  which  implies  a  much  larger  molecule  than  that  of 
lecithin.  It  is  made  up  of  glycerophosphoric  acid,  three  fatty  acids 
and  a  nitrogen-containing  base  that  is  not  choline.  The  fatty  acids 
are  mainly  highly  unsaturated  and  are  thought  to  belong  to  the  lino- 
leic  or  the  linolenic  acid  series.  Cuorin  is  even  more  autooxidizable 


PHOSPHORUS  METABOLISM 


69 


than  lecithin,  and  the  properties  are  much  altered  by  such  oxida- 
tion. MacLean  (1912a,  1912b)  finds  cuorin  in  horse  kidney  and 
muscle. 

OTHER   MONAMINO-DIPHOSPHATIDS 

Other  monamino-diphosphatids  similar  to  cuorin,  but  not  iden- 
tical with  it,  have  been  obtained  by  Baskoff  (1908)  from  horse  liver, 
and  by  MacLean  (1908b,  1909b)  from  egg-yolk.  Elementary  analy- 
ses of  the  three  substances  are  given  here : 

ELEMENTARY   ANALYSES    OF    MONAMINO-DIPHOSPHATIDS— Percent 


Author 

Date  and 
Refer- 
ence 

Source 

C 

H 

N 

P 

O 

S 

N:P 

Erlandsen 

Baskoff   . . 
MacLean  . 

1906, 
1907 
1908 
1909b 

Heart 

Liver 
Egg-yolk 

61.63 

61.12 
59.12 

9.03 

8.95 
9.44 

1.015 

1.23 

0.812 

4.46 

4.00 
3.59 

23.86 

(24.04) 
27.048 

None 
0.6 

1:1.99 

1:1.47 
1:2 

Baskoff  called  his  compound  "heparphosphatid."  It  contained 
sulphur.  The  N-component  was  thought  to  be  other  than  choline, 
but  was  undetermined.  It  had  a  lower  fatty  acid  content  than  leci- 
thin. It  did  not  react  with  sugar.  MacLean  found  his  compound 
in  about  equal  amount  with  cephalin  in  egg-yolk. 

Ill      TRIAMINO-DIPHOSPHATIDS 

Triamino-diphosphatids  have  been  obtained  only  by  Frankel 
and  his  co-workers  as  CdCl2  compounds  (Frankel  and  Nogueira, 
1909a,  1909b;  Frankel  and  Linnert,  1910).  The  methods  used  are 
unreliable.  These  compounds  were  obtained  from  kidney  and  from 
brain.  Only  that  from  the  brain,  which  is  called  "sahidin"  has  been 
studied  in  detail.  From  hydrolysis  it  appears  to  be  a  compound  of 
choline  with  glycerophosphoric  acid  and  fatty  acids,  the  latter  being 
of  both  saturated  and  unsaturated  type.  It  is  quite  conceivable 
that  the  compound  under  examination  was  a  monamino-monophos- 
phatid  of  the  lecithin  type  containing  a  nitrogenous  substance  as  an 
impurity. 

B.     SATURATED  PHOSPHATIDS 

I.    DIAMINO-MONOPHOSPHATIDS 

This  class  of  compounds  appear  to  be  primary  constituents  of 
cells,  but  they  have  not  been  studied  sufficiently  to  furnish  definite 
conceptions  as  to  their  constitution  or  their  relations  to  one  another. 


70  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL,  5 

SPHINGOMYELIN 

Thudichum,  and  Rosenheim  and  Tebb  (Rosenheim  and  Tebb, 
1907,  1908a,  1908b,  1908c,  1909a,  1909c,  1910)  have  felt  that  they 
showed  the  so-called  "protagon"  of  the  brain  to  be  mainly  a  mixture 
of  a  diamino-monophosphatid,  which  Thudichum  named  sphingo- 
myelin, and  cerebrosides.  Rosenheim  and  Tebb  found  the  same 
characteristics  in  the  substance  isolated  from  the  cortex  of  the  ad- 
renals, but  with  a  smaller  amount  of  sphingomyelin.  Thudichum 
gives  the  formula  C52H104N2PO9+H2O  for  sphingomyelin.  Rosen- 
heim and  Tebb  (1908c)  report  an  elementary  analysis  with  3.46 
percent  P,  from  which  Bang  computes  the  formula  C49Hlo4N2PO10. 
If  the  work  of  these  two  sets  of  observations  is  correct,  sphing- 
omyelin is,  with  carnaubon  only,  an  exception  among  the  phos- 
phatids  in  that  it  contains  no  glycerin,  and  therefore  is  not  a 
glycerophosphoric  acid  compound.  It  does  show  phosphoric 
acid,  fatty  acids  (perhaps  only  one,  which  Thudichum  iden- 
tified as  an  isomer  of  ordinary  stearic  acid)  and  choline 
(or  neurine)  and  a  crystalline  alcohol.  Rosenheim  and  Tebb  (1908b) 
made  a  study  of  the  peculiar  property  of  sphingomyelin  by  which  it 
separates  from  its  pyridine  solution,  or  a  pyridine  solution  of  pro- 
tagon, in  doubly  refracting  spheroid  crystals  which  are  strongly 
laevorotatory,  the  dextrorotation  of  the  original  protagon  solution 
thus  being  gradually  lessened  and  finally  changed  to  laevorotation. 
Since  this  substance  is  probably  widely  distributed  in  cellular  tis- 
sues, it  is  desirable  that  it  should  receive  much  further  study. 

OTHER  DIAMINO-MONOPHOSPHATIDS 

Other  diamino-monophosphatids  are  the  apomyelin  and  amido- 
myelin  of  Thudichum,  a  phosphatid  isolated  from  liver  by  Baskoff 
(1908)  which  showed  the  ratio  P:N=1 :2.55,  one  taken  from  kidney 
by  Nogueira  for  which  the  formula  C34H72N2PO10  is  given,  and  two 
others  the  properties  of  which  are  reported  as  resembling  those  of 
sphingomyelin.  Of  these,  that  of  Stern  and  Thierf elder  (1907), 
was  obtained  from  egg-yolk,  and  the  elementary  analysis  showed 
3.22  percent  P  and  a  ratio  P:N— 1:1.9.  One  of  the  compounds 
which  Erlandsen  obtained  as  a  CdCl2  combination,  but  was  not  able 
to  isolate  free,  seemed  to  have  the  formula  C40H75N2PO12.  It  was  ob- 
tained from  both  types  of  muscle,  though  apparently  it  was  not 
present  in  the  free  state  but  in  protein  combination.  Cleavage  in- 
dicated but  one  molecule  of  fatty  acid  and  probably  two  basic 
radicals.  The  fatty  acid  was  an  oxyacid.  Otherwise  it  much  re- 
sembled lecithin. 


PHOSPHORUS  METABOLISM  71 

Burow  (1910)  finds  three  iron-containing  phosphatids  in  the 
spleen  of  cattle  and  of  man.  Only  one  of  these  was  obtained  in  suf- 
ficient amount  for  analysis  and  that  proved  to  be  a  diamino-mono- 
phosphatid.  The  name  "ferroid"  was  proposed  to  show  its  iron- 
lipoid  character. 

Baskoff's  (1908)  analyses  of  purified  jecorin  indicate  that  that 
compound  may  be  a  diamino-monophosphatid  (see  p.  65). 

II.    TRIAMINO-MONOPHOSPHATIDS 

The  triamino-monophosphatids  which  have  been  reported  are 
the  neottin  of  egg-yolk,  found  by  Frankel  and  Bolaffio  (1908),  and 
the  carnaubon  of  kidney,  found  by  Dunham  and  Jacobson  (1910). 

NEOTTIN 

Neottin  is  a  saturated  compound,  optically  inactive.  It  is 
thought  to  contain  choline  and  three  saturated  fatty  acids,  stearic, 
probably  palmitic  and  perhaps  cerebronic. 

CARNAUBON 

Carnaubon  differs  decidedly  from  all  other  animal  phosphatids 
that  have  been  examined,  except  jecorin,  in  that  it  contains  a  sugar 
(galactose  or  aminogalactose)  which  is  but  incompletely  split  off  in 
simple  hydrolysis  and  therefore  must  form  an  essential  part  of  the 
phosphatid  molecule.  Such  compounds  are  common  in  plants.  No 
glycerin  is  found  in  carnaubon,  and  Dunham  and  Jacobson  suggest 
that  the  structure  may  be  similar  to  that  of  lecithin,  but  with, the 
sugar  in  the  place  of  glycerin.  Three  fatty  acids  were  found, 
carnaubic,  stearic  and  palmitic,  with  phosphoric  acid  and  choline. 
Assuming  that  carnaubon  is  made  up  of  aminogalactose,  these  three 
fatty  acids  and  one  phosphoric  acid  in  combination  with  two  choline 
groups,  the  formula  C74H150N3PO13  is  proposed. 

MacLean  (1912a,  1912-13)  isolated  from  horse  kidney  a  sub- 
stance having  all  the  properties  of  this  compound  of  Dunham  and 
Jacobson,  but  it  was  a  diamino-monophosphatid.  He  thinks  it  prob- 
able that  carnaubon  is  not  a  tri-,  but  a  diamino-monophosphatid  and 
that  the  methods  used  by  Dunham  and  Jacobson  were  inefficient  to 
obtain  a  pure  substance.  By  water  extractions  MacLean  (1912a, 
1912b)  separates  water-soluble  substances  which  contain  but  little 
phosphorus,  and  are  not  of  the  nature  of  ordinary  phosphatids,  but 
are  easily  confused  with  them  in  the  processes  ordinarily  used  for 
isolation  of  phosphatids.  As  these  substances  have  a  high  nitrogen 
content,  it  may  be  that  several  of  the  substances  hitherto  described 
as  containing  large  percentages  of  nitrogen  are  really  mixtures  of 


72  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

simpler  bodies  with  these.      He  recommends  a  purification  by  a  pro- 
cess of  emulsification  with  water  and  precipitation  with  acetone. 

III.    PROTAGON 

For  a  long  time  the  name  "protagon"  was  in  use  for  a  substance 
isolated  from  brain  and  thought  to  be  the  principal,  perhaps  the 
only,  phosphatid  of  brain.  It  was  found  also  in  many  other  parts 
of  the  body.  That  these  preparations  were  crystalline  and  fairly 
constant  in  composition  even  after  recrystallization  justified  the 
belief  that  protagon  was  a  chemical  unit ;  later  work,  however,  has 
shown  that  such  a  conception  is  untenable.  Since  the  first  finding 
and  naming  of  protagon  by  Liebreich  in  1865  it  has  received  much 
attention.      A  list  of  references  is  given  below. 

The  question  of  protagon  being  the  main  or  only  constituent  of 
the  brain  was  satisfactorily  answered  by  Frizzell's  analyses,  report- 
ed by  Chittenden,  and  especially  by  the  abundant  evidence  from 
Thudichum's  work.  Thudichum  also  showed  that  it  is  a  mixture, 
and  that  it  does  not  contain  lecithin.  Thudichum's  work,  however, 
seems  not  to  have  met  with  due  recognition  until  after  his  death. 
Further  evidence  against  the  chemical  entity  of  protagon  has  been 
produced  mainly  by  Gies  and  his  associates,  and  by  Rosenheim  and 
Tebb.  They  have  shown  that  it  is  made  up  of  phosphorus-rich  and 
phosphorus-poor  components  mixed  in  variable  proportions.  Kossel 
and  Freytag  had  already  recognized  the  presence  of  cerebrosides, 
which  are  phosphorus-free  compounds  consisting  of  the  sugar  galac- 
tose, fatty  acids  and  a  nitrogen  complex.  Thierf elder  and  his  as- 
sociates have  separated  and  studied  cerebron,  another  phosphorus- 
free  compound.  The  phosphorus  compound  was  identified  by 
Thudichum  and  by  Rosenheim  and  Tebb,  as  sphingomyelin,  but  it  is 
perhaps  not  a  single  phosphatid.  Sulphur  has  usually  been  found 
in  protagon,  and  apparently  in  organic  combination,  and  Koch 
thought  that  he  had  evidence  of  a  complex  containing  an  ethereal 
sulphuric  acid  combined  with  a  phosphatid  and  a  cerebroside,  as  one 
of  the  substances  present  in  the  mixture.  Frankel  concludes  that 
protagon  is  a  mixture  of  members  of  the  group  which  he  designates 
"galacto-phospho-sulphatids,"  or  combinations  of  phosphatids  and 
sulphatids  with  galactose.  It  is  not  to  be  looked  upon  as  a  chemical 
unit. 

References  on  protagon:  Liebreich,  1865;  Diaconow,  1867a; 
Gamgee  and  Blankenhorn,  1879a,  1879b,  1879c,  1880;  Baumstark, 
1885;  A.  Kossel,  1891b;  A.  Kossel  and  Freytag,  1893;  Ruppel,  1895; 
Chittenden,  1897 ;  Noll,  1899 ;  Worner  and  Thierf  elder,  1900 ;  Kita- 


PHOSPHORUS  METABOLISM  73 

gawa  and  Thierf  elder,  1906 ;  Ulpiani  and  Lelli,  1902 ;  Barbieri,  1905 ; 
Lesem  and  Gies*,  1903 ;  Cramer,  1904 ;  Orgler,  1904 ;  Posner  and  Gies, 
1905;  Lochhead  and  Cramer,  1907;  Gies,  1907;  Steel  and  Gies, 
1907b ;  Wilson  and  Cramer,  1908 ;  Cohen  and  Gies,  1908 ;  Rosenheim 
and  Tebb,  1907,  1908a,  1908b,  1908c,  1909a,  1909c,  1910;  W.  Koch, 
1907b,  1910a,  1912 ;  Frankel,  1908. 

D.    PLANT  PHOSPHATIDS 

Apparently  phosphatids  are  as  widely  distributed  (and  there- 
fore probably  essential)  in  plant  cells  as  in  animal  cells,  and  are  as 
varied  in  their  make-up,  though  the  work  of  differentiation  and 
classification  has  not  yet  been  carried  even  as  far  as  it  has  for  those 
of  animal  origin.  Nearly  all  of  the  investigations  we  have  found 
reported  have  been  made  by  E.  Schulze  and  E.  Winterstein  and  their 
associates  (Schulze  and  Steiger,  1889 ;  Schulze  and  Likiernik,  1891a, 
1891b;  Schulze  and  Frankfurt,  1894;  Schulze,  1895,  1897,  1907, 
1908a,  1908b;  Schulze  and  Winterstein,  1903;  Hiestand,  1906;  Win- 
terstein and  Hiestand,  1906,  1908 ;  Winterstein  and  Stegmann, 
1909a,  1909b;  Winterstein  and  K.  Smolenski,  1909;  K.  Smolenski, 
1909;  Schulze  and  Pfenninger,  1911;  Trier,  1911,  1913a,  1913b, 
1913c,  1913d).  A  few  other  articles  should  be  mentioned,  especially 
that  of  Njegovan  (1911) ;  also  those  of  Wintgen  and  Keller  (1906) ; 
Parrozzani  (1909);  Vorbrodt  (1910)  and  Bernardini  (1912). 

It  was  at  first  supposed  that  all  the  phosphorus-containing  fat- 
like substances  obtained  were  identical  with  those  of  animal  tissues, 
and,  as  for  the  latter,  so  for  these,  the  name  lecithin  was  used,  and 
the  cleavage  products  somewhat  supported  the  idea ;  but  it  was  later 
shown  that  there  are  several  different  phosphatids  here  and  that  the 
true  lecithins  are  not  found  at  all  in  plants.  The  phosphatids  of 
the  two  kingdoms  are  essentially  different,  in  that  vegetable  phos- 
phatids nearly  or  quite  always  contain  a  sugar,  apparently  firmly 
bound  and  in  constant  stoichiometric  relation  with  the  rest  of  the 
molecule,  such  as  is  not  the  case  in  animal  phosphatids,  with  the 
probable  exception  of  jecorin,  and  perhaps  carnaubon.  The  amount 
of  carbohydrate  found  has  been  widely  different  in  different  prepar- 
ations, suggesting  that  a  part,  at  least,  of  it  is  present  as  a  phos- 
phatid-sugar  combination  such  as  the  lecithin-sugars  mentioned 
above.  The  plant  phosphatids  contain,  together  with  the  sugar, 
glycerophosphoric  acid,  fatty  acids  and  choline,  and  sometimes  other 
nitrogenous  residues,  either  basic  or  amino  acid. 


74  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

ARTIFICIAL  SYNTHESIS  OF  PHOSPHATIDS 

Glycerophosphates  have  been  made  by  several  investigators, 
and  they  have  found  extensive  use  as  drugs.  See  Adrian 
and  Trillet  (1897,  1898),  Lumiere,  A.  L.,  Lumiere  and  F.  Perrin 
(1901),  Carre  (1904),  Willstatter  and  Liidecke  (1904),  Power  and 
Tutin  (1905),  Tutin  and  Hann  (1906),  Neuberg  and  Kretschmer 
(1911),  Langheld  (1910,  1911,  1912),  DuBois  (1914),  and  also  a  re- 
view of  literature  by  Merck  (1911). 

The  question  of  the  identity  of  the  synthetic  products  with  the 
glycerophosphate  of  lecithin  was  considered  by  Willstatter  and 
Liidecke,  Tutin  and  Hann,  and  Power  and  Tutin,  with  evidence  that 
they  are  not  identical,  and  that  the  difference  is  in  the  structure  of 
the  molecule.  Power  and  Tutin  say  that,  in  the  substances  handled, 
the  mono-ester,  C3H5  (OH)  2O.PO  (OH)  2,  has  often  been  contaminated 

A 

with  the  di-ester,  C3H5OH  ^POCOH).  Tutin  and  Hann  con- 
clude that  both  the  synthetic  and  the  natural  products  are  mixtures 
of  the  asymmetric  or  a-acid,  CH2O.P03H2, 

CH.OH 

CH2.OH 
and  the  symmetrical  or  /?-acid,  CH2.OH 

CH.O.P03H2 

CH2.OH 

DuBois  (1914)  discusses  these  and  other  possible  glycerophos- 
phates, and  gives  in  detail  the  properties  of  such  as  are  known.  Con- 
sidering the  evidence  brought  forth  by  others  in  comparing  the  nat- 
ural and  synthetical  products  leads  him  to  the  "conclusion  that  the 
synthetical  glycerophosphates  produced  at  low  temperatures  (100- 
110°),  and  the  natural  glycerophosphates  obtained  from  lecithin  are 
almost  identical,  and  in  all  probability,  a  mixture  of  the  a-  and  /?- 
isomerides,  in  which  the  a-isomeride  predominates." 

According  to  Francois  and  Boismenu  (1913)  commercial  cal- 
cium glycerophosphate  may  be  a  mixture  of  salts  of  5  forms  of  the 
acid,  namely  the  tri-ether,  and  the  a-  and  /?-  forms  of  both  di-  and 
mono-ethers.  The  calcium  salt  of  the  tri-ether,  however,  is  so  diffi- 
cultly soluble  that  it  is  not  usually  permitted  by  the  manufacturer  to 
be  present  in  the  commercial  article. 


PHOSPHORUS  METABOLISM  75 

THE  PHOSPHORUS  OF  PHOSPHATIDS 

In  the  phosphatids,  and  in  no  other  natural  products,  so  far  as 
known,  phosphorus  is  present  as  glycerophosphoric  acid,  a  glycerin 
ester  of  orthophosphoric  acid.  The  most  ready  cleavage  sets  free  this 
glycerophosphoric  acid,  and  further  cleavage  produces  free  ortho- 
phosphoric  acid.      This  relation  seems  to  be  unquestioned. 

See  Imbert  and  Belugon  (1897),  with  regard  to  the  reactions  of 
glycerophosphoric  and  phosphoric  acids  with  bases,  and  Malengreau 
and  Prigent  (1911)  with  regard  to  the  hydrolysis  of  glycerophos- 
phoric acid. 

For  general  discussions  on  the  phosphatids  see  Thudichum  (1901), 
Hiestand  (1906),  Bang  (1909),  Frankel  (1908,  1909a),  MacLean  (1909d),  Bang 
(1911a)  and  Merck  (1911,  1912). 


76  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  6 


PART  II. 
THE  PHOSPHORUS  COMPOUNDS  OF  FOODS 

GENERAL  DISCUSSION 

A  statement  of  the  phosphorus  content  of  a  foodstuff  is  of 
slight  value  apart  from  a  consideration  of  other  nutrient  constitu- 
ents, but  we  can  not,  within  the  scope  of  this  article,  go  into  the 
matter  of  general  composition  of  foods.  In  our  selection  of  material 
for  presentation  in  this  section,  therefore,  we  have  sought  to  include 
especially  data  showing  the  connection  of  the  phosphorus  content  of 
foods  with  other  matters  of  interest,  particularly  as  related  to  gen- 
eral type  of  food  products,  or  to  conditions  of  growth,  preparation 
or  manufacture ;  and  also  such  material  as  is  available  showing  the 
differential  estimation  of  the  various  groups  or  kinds  of  phosphorus 
compounds  in  foodstuffs.  For  data  on  total  phosphorus  of  foods  see 
the  works  cited  on  p.  103. 

Analyses  of  meats,  milks,  and  eggs  will  be  found  in  the  section 
or  Phosphorus  of  Animal  Bodies  and  Products,  and  are  not  included 
here  except  as  they  appear  in  miscellaneous  tables. 

From  the  following  tables  of  Forbes,  Beegle  and  Mensching 
(1913)  we  note  that  among  the  milling  products  of  wheat  the  phos- 
phorus is  contained  principally  in  the  byproducts  which  are  used  as 
foods  for  live-stock.  The  white  flour  is  that  portion  of  the  grain 
of  wheat  which  contains  the  least  phosphorus.  Wheat  gluten,  such 
as  is  used  as  a  diabetic  flour  is  also  poor  in  phosphorus.  The  bran 
contains  more  phosphorus  than  any  other  part  of  the  kernel. 

For  further  details  regarding  the  phosphorus  constituents  of 
wheat  see  Osborne  and  his  associates,  p.  91,  Rengniez,  p.  78, 
Rising,  p.  79,  Ames  and  Boltz,  p.  96-98,  and  Swanson,  p.  80, 
of  this  work;  also  Girard  (1884),  Girard  and  Lindet  (1903),  Balland 
(1903),  Surmont  and  Dehon  (1903,  1904)  and  Fauvel  (1907)  ;  also 
references  on  p.  103  to  general  works  on  food  analysis  and  nutri- 
tion. 


PHOSPHOEUS  METABOLISM 


77 


Corn  contains  less  phosphorus  than  do  oats  and  wheat,  the  pearl 
hominy  and  bolted  corn  meal  used  by  human  beings  containing  less 
phosphorus  than  the  whole  grain.  Gluten  feed  contains  more 
phosphorus  than  does  the  whole  corn,  and  distiller's  grains  about 
the  same  as  corn. 

•  Kafir  corn  contains  less  phosphorus  than  does  Indian  corn, 
while  polished  rice  is  very  poor  in  phosphorus,  about  the  same  as 
pearl  hominy.  Rice  polish,  however,  which  is  fed  to  live-stock,  is 
exceedingly  rich  in  phosphorus. 

On  a  dry  basis  the  fruits  are  very  low  in  phosphorus ;  such  veg- 
etables as  onions,  cabbage,  beets,  and  potatoes  contain  much  more. 

Among  various  kinds  of  roughage,  leguminous  forage  contains 
considerable  phosphorus,  as  also  does  blue-grass.  Timothy  hay, 
corn  stover  and  wheat  straw  contain  much  less.  The  phosphorus 
content  of  all  sorts  of  roughage  is  much  affected  by  the  soil. 

Leguminous  seeds  are  characteristically  rich  in  phosphorus, 
much  richer  than  the  leaves  and  stems  of  the  same  species, 
while  the  oil  meals  are  still  richer,  cottonseed  meal  excelling  linseed 
oil  meal  in  this  regard. 

MINERAL  ELEMENTS  OF  CEREAL  PRODUCTS— Parts  per  100  of  Dry 

Substance  (Forbes,  Beegle  and  Mensching,  1913) 


Potas- 
sium 

Sodium 

Cal- 
cium 

Magne- 
sium 

Sul- 
phur 

Chlor- 
ine 

Phos- 
phorus 

Inor- 
ganic 
phos- 
phorus 

Organ- 
ic 
phos- 
phorus 

Wheat 

- 

.590 
.058' 
.156 
1.464 
1.147 
.323 
.007 
.425 
.396 
.192 
.410 
.153 
.272 
.014 
.045 
.185 
.219 
.460 
.288 
.040 
1.279 

.035 
.127 
.583 
.223 
.186 
.788 
.031 
.733 
.030 
.113 
.000 
.000 
.461 
.154 
.077 
.278 
1.458 
.184 
.066 
.032 
.124 

.056 
.022 
.038 
.139 
.108 
.078 
.085 
.134 
.014 
.015 
.030 
.005 
.268 
.047 
.142 
.169 
.159 
.112 
.013 
.009 
.030 

.142 
.019 
.004 
.590 
.430 
.372 
.049 
.324 
.126 
.122 
.088 
.036 
.239 
.054 
.195 
.   .172 
.194 
.130 
.142 
.028 
.741 

.224 
.168 
.198 
.297 
.263 
.355 
1.000 
.285 
.171 
.122 
.124 
.182 
.636 
.509 
.408 
.419 
.864 
.214 
.186 
.114 
.189 

.095 
.081 
.958 
1.000 
.029 
.077 
.055 
.156 
.073 
.070 
.052 
.052 
.098 
.065 
.028 
.062 
.389 
.077 
.117 
.040 
.151 

.425 
.102 
.135 

1.233 
.984 

1.147 
.220 
.928 
.303 
.264 
.156 
.111 
.589 
.314 
.458 
.503 
.746 
.434 
.271 
.104 

1.684 

.038 

.017 
.043 
.034 
.069 

.037 
.098 
.028 
.019 
.031 
.019 
.106 
.056 
.018 
.162 
.471 
.059 
.012 
.003 
.028 

.387 

.085 

White   bread    

Wheat  bran    .  . 

Wheat  middlings   .  .*  .  .  . 

.092 

1.199 

.915 

Wheat    gluten    

Red  dog  flour    

.183 

,  .830 

.275 

.245 
.125 

Pearl    hominy    

.092 
.483 

Distiller's  grains,  corn.. 
Distiller's  grains,  rye   .  . 

.258 
.440 
.341 
.275 

.375 
.259 

.101 

1.656 

The  animal  products  are,  on  a  dry  basis,  rich  in  phosphorus, 
even  whey  containing  notable  amounts,  while,  of  course,  such  foods 
as  tankage,  which  contain  some  bone,  are  much  richer  than  any  oth- 
er foods  in  phosphorus.  Milk  and  eggs  do  not  differ  greatly  in  phos- 
phorus content.  Both  are  rich  in  phosphorus,  and  contain  much 
more  of  this  element  than  does  meat. 


78 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


MINERAL  ELEMENTS  OF  FRUITS,  VEGETABLES  AND  ROUGHAGE— 

Parts  per  100  of  Dry  Substance  (Forbes,  Beegle  and 

Mensching,  1913) 


Potas- 
sium 

Sodium 

Cal- 
cium 

Magne- 
sium 

Sul- 
phur 

Chlor- 
ine 

Phos- 
phorus 

Inor- 
ganic 
phos- 
phorus 

Organ- 
ic 
phos- 
phorus 

.802 

1.347 

1.291 

.667 

1.442 

2.484 

1.208 

1.547 

3.870 

.347 

1.840 

1.774 

.873 

.832 

.613 

1.338 

1.847 

1.405 

.842 

.132 

.066 
.045 
.240 
.115 
.097 
.028 
.061 
.175 
.714 
.185 
.067 
.145 
.722 
.489 
.345 
.099 
.065 
.141 
.237 
.135 

.027 
.076 
.037 
.084 
.261 
.590 
.084 
.027 
.131 
.729 
1.236 
1.378 
2.029 
1.130 
.192 
.326 
.507 
.336 
.217 
.780 

.033 
.056 
.129 
.086 
.136 
.209 
.215 
.331 
.358 
.283 
.292 
.692 
1.096 
.400 
.111 
.262 
.092 
.240 
.063 
.570 

.044 
.066 
.021 
.087 
.601 
.901 
.117 
.141 
.224 
.138 
.190 
.259 
.352 
.298 
.162 
.159 
.187 
.334 
.159 
2.090 

.037 
.050 
.421 
.285 
.183 
.243 
.069 
.055 

1.380 
.048 
.259 
.084 
.167 
.161 
.199 

1.230 
.308 
.234 
.209 
.040 

.064 
.110 
.119* 
.077 
.323 
.262 
.186 
.270 
.260 
.069 
.183 
.237 
.283 
.238 
.123 
.173 
.102 
.242 
.038 
.024 

.033 

.098 
.089 
.037 
.210 
.136 
.138 
.130 
.174 
.006 
.080 
.121 
.152 
.122 
.052 
.072 
.039 
.142 
.015 
.003 

.031 

.012 

.030 

Date    

.040 

.113 

.126 

.048 

.140 

.086 
.063 

.103 

.116 

.131  ' 

.116 

.071 

.101 

.063 

.100 
.023 
.021 

MINERAL  ELEMENTS  OF  LEGUMINOUS  SEEDS,  NITROGENOUS  CON- 
CENTRATES  AND   ANIMAL   PRODUCTS— Parts   per   100   of   Dry 
Substance  (Forbes,  Beegle  and  Mensching,  1913) 


Soy  beans    , 

Navy  beans .  , 

Cowpeas    

Peanuts     

Linseed  oil  meal 
Cottonseed  meal 

Milk,  skim 

Whey    , 

Mutton    

Eggs    

Tankage    

"Banner"  bone  flour 

Blood,   swine    

"Black    albumen"     . 


Potas- 
sium 


2.095 

1.390 

1.636 

.061 

1.224 

1.811 

1.272 

2.762 

.624 

.206 

.601 

.065 

1.040 

.027 


Sodium 


.380 

.086 

.189 

.563 

.282 

.283 

.488 

.459 

.214 

.389 

1.830 

.091 

1.370 

1.247 


Cal- 
cium 


Magne- 
sium 


.230 
.235 
.117 
.068 
.403 
.291 

1.336 
.721 
.006 
.250 

3.242 

23.990 

.031 

.039 


.244 
.206 
.243 
.180 
.544 
.599 
.146 
.138 
.062 
.059 
.159 
1.160 
.028 
.011 


Sul- 
phur 


.444 
.224 
.280 
.254 
.455 
.536 
.357 
.139 
.607 
.762 
.669 

.647 
.820 


Chlor- 
ine 


.025 
.047 
.047 
.024 
.095 
.042 
.953 

1.948 
.235 
.621 

2.687 

1.200 
1.550 


Phos- 
phorus 


.649 
.429 
.532 
.399 
.786 

1.479 
.979 
.640 
.474 
.856 

1.789 

14.940 

.280 

.122 


Inor- 
ganic 
phos- 
phorus 


.017 
.088 
.023 
.049 

.078 
.551 
.402 
.230 
trace 

14.940 
.076 
.037 


Organ- 
ic 

phos- 
phorus 


.632 
.341 
.509 
.350 

1.401 
.428 
.238 
.244 
.856 


.204 
.085 


From  the  work  of  Vorbrodt  (1910)  we  quote  the  table  on  the 
following  page  setting  forth  a  separation  of  general  groups  of  phos- 
phorus compounds. 

Rengniez  (1911)  reports  most  of  the  phosphorus  of  flour  to  be 
in  the  forms  of  phytin  and  nuclein.  The  highest  phosphorus  con- 
tent was  found  in  flour  made  from  the  germs.  Wheat  germ  flour 
showed  2.9  percent  total  phosphoric  acid,  of  which  2.17  percent  was 
in  the  form  of  phytin  and  0.70  percent  in  the  form  of  nuclein. 


PHOSPHORUS  METABOLISM 


79 


CONTENT  OF  DIFFERENT  FORMS  OF  PHOSPHORIC  ACID  IN  SEEDS 

Vorbrodt,  (1910) 


Seeds  of 


Indian  corn  (Zea  Mays)  .  . . 

Wheat  (Triticum  sativum) 

Barley (Hordeum  distich.)! 

"II 

Rye  (Secale  cereale) 

Lentil  (Lens  esculenta)    II 
Windsor  bean  (  Vicia  faba 

minor) 

Rape  (Brassica  napus  olei- 

fera) 

Hemp  (  Cannabis  sativa)  . . 

Spruce  (Picea  excelsa) 

Pine  (Pinus  cembrd) 


Total 
P2O5 

Percent 


0.814 

1.04 

1.07 

0.83 

0.98 

0.70 

1.07 

1.23 

1.74 
1.57 
1.07 


Mineral  P2O5 


Percent  Percent 
of  seed  of  total 


0.050 

0.134 

0.15 

0.127 

0.302 

0.134 

0.081 

0.102 
0.213 
0.138 
0.079 


6.14 
12.88 
14.02 
15.30 
30.82 
19.14 

7.57 

8.29 

12.24 

8.70 

7.38 


Soluble  organic 
P2O5 


Percent 
of  seed 


0.398 
0.311 
0.389 
0.285 
0.283 
0.065 

0.047 

0.468 
0.261 
0.340 
0.154 


Percent 
of  total 


29.90 
36.36 
34.34 
28.88 
9.29 

4.39 

38.05 
15.00 
21.65 
14.39 


Protein  P2O5 


Percent 
of  seed 


0.319 
0.567 
0.531 
0.388 
0.382 
0.435 

0.894 

0.66 
1.266 
1.092 
0.837 


Percent 
of  total 


39.18 
54.51 
49.63 
46.75 
38.98 
61.71 

83.56 

53.66 
72.76 
69.56 

78.22 


Lecithin  P2O5 


Percent 
of  seed 


0.047 
0.028 


0.030 
0.013 
0.069 

0.048 


0.029 
0.053 


Percent 
of  total 


5.77 
2.69 

1.33 
9.86 

4.49 


1.85 
4.95 


General  phosphorus  separations  are  also  quoted  from  the  work 
of  Rising  (1910). 


PHOSPHORUS  COMPOUNDS  IN  PEAS,   BEANS  AND  FLOURS 

Rising  (1910) 


Phos.  of  nu- 

Total 

Phosphorus  of 

Phosphorus  of 

Phosphorus 

clein  sub- 
stance and 
phosphopro- 
tein 

Mois- 
ture 

phosphorus 

phosphatids 

phosphates 

of  phytin 

Sum   of 

Per- 

Per- 

Per- 

Per- 

Per- 

of solids 

other 

Per- 

cent 

Per- 

cent 

Per- 

cent 

Per- 

cent 

cent 

found 

deter- 

cent 

of  total 

cent 

of  total 

cent 

of 

cent 

of 

directly 

mina- 
tions 

P 

P 

total 
P 

total 
P 

Yellow  peas 

13.7 

0.579 

0.565 

0.0586 

10.1 

0.019 

3.3 

0.110 

19 

0.378 

65.1 

Brown  beans.. 

11.8 

0.560 

0.5167 

0.0397 

7. 

0.019 

3.4 

0.290 

52 

0.168 

30. 

Rye  flour 

11.85 

0.313 

0.3278 

0.0073 

2.8 

0.090 

2.8  1 

0.0885 

25 

0.142 

45.3 

13.5 

0.1665 

0.-1891 

0.0067 

4. 

0.0116 

7. 

0.1144 

69 

0.0564 

33. 

Graham  flour. . 

12.4 

0.287 

0.2400 

0.0058 

2. 

0.0381 

13.2 

0.0819 

29 

0.1142 

39. 

(1)      Apparently  a  misprint  for  28. 

Hooper  (1911)  states  that  unmilled  rice  contains  about  0.65 
percent  P205,  while  polished  rice  contains  0.38  percent.  The  por- 
tion of  the  rice  which  is  removed  by  the  milling  process  contains 
3.36  percent  P205.  He  reports  Indian  wheat  to  contain  0.69  percent 
P205,  and  flour  from  the  same  0.21  percent  P205. 

Aron  and  Hocson  (1911a)  found  in  husked  rice  0.7-0.8  percent 
P205;  undermilled  rice,  0.45-0.6  percent;  and  overmilled  rice,  0.15- 
0.35  percent. 

Bernardini  (1912)  gives  the  distribution  of  phosphorus  in  rice 
as  follows : 


80  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

DISTRIBUTION  OF  PHOSPHORUS  IN  RICE  (Bernardini,  1912) 


P2O5  contained  in 

In  embryo,  percent 

In  whole  seed,   percent 

Total    

6.20 
0.04 
0.22 
5.14 
0.04 
0.76 

0.950 

0.003 

0.018 

Phytin    

0.436 

0.502 

Adler  (1912a,  1912b)  investigated  the  nature  of  the  phospho- 
rus compounds  of  brewing  products.  Eighty  percent  of  the  phos- 
phorus of  beer  is  inorganic;  20  percent  organic.  Of  the  organic 
phosphorus  a  part  is  combined  with  carbohydrates,  and  a  part  with 
proteins.  Lecithin  and  phytin  are  not  present.  Of  the  phos- 
phates, half  are  united  with  alkalis  and  half  with  alkaline  earths. 
Wort  contains  both  organic  and  inorganic  phosphorus,  and  in  about 
the  same  proportion  as  in  beer.  Malt  contains  alkali  and  alkaline 
earth  phosphates,  as  well  as  phytin.  Malt  sprouts  also  contain  al- 
kali and  alkaline  earth  phosphates,  in  greater  amounts  than  either 
the  barley  or  the  malt. 

Swanson  (1912)  reported  investigations  with  regard  to  the  re- 
lations between  the  percentages  of  acidity  and  percentages  of  ash, 
amino  compounds,  and  total  and  water-soluble  phosphorus  of  wheat 
flours,  the  work  being  part  of  a  general  study  on  the  relation  be- 
tween chemical  composition  and  baking  qualities  of  flour.  The 
total  phosphorus  varies  from  0.093  percent  (in  second  middlings) 
to  0.373  percent  (in  ship-duster  flour),  the  amount  in  wheat  being 
given  as  0.482  percent;  the  water-soluble  phosphorus  at  25°  varies 
from  0.017  percent  (first  and  second  middlings)  to  0.114  percent 
(fifth  break) ;  and  the  water-soluble  at  40°,  from  0.023  percent 
(first  middlings)  to  0.191  percent  (ship-duster  flour),  that  of  wheat 
being  0.218  percent. 

ORGANIC  AND  INORGANIC  PHOSPHORUS  IN  FOODS 

Methods  for  the  estimation  of  inorganic  phosphorus  as  distinct 
from  organic  phosphorus  in  foods  are  not  yet  sufficiently  well  estab- 
lished to  warrant  the  placing  of  emphasis  upon  minor  differences  in 
results  obtained.  One  of  us  (E.  B.  F.)  in  collaboration  with  Leh- 
mann,  Collison  and  Whittier  published  methods  for  this  estimation 
in  both  plant  and  animal  tissues  (Ohio  Agr.  Exp.  Sta.  Bui.  215),  and 
results  obtained  by  these  methods  are  submitted  in  the  tables  on  pp. 
77  and  78.      In  later  studies  of  these  methods  the  one  used  for 


PHOSPHORUS  METABOLISM 


81 


animal  tissues  has  promised  well,  while  the  outcome  of  further  work 
with  the  method  for  vegetable  products  seems  problematical.  Cer- 
tain general  conclusions,  however,  we  feel  it  safe  to  draw  from  these 
data.  Thus,  all  of  the  parts  of  plants  contain  inorganic  as  well  as 
organic  phosphorus.  The  proportion  of  the  total  phosphorus  which 
is  inorganic  is,  in  general,  much  higher  in  leaves,  stems,  fruits  and 
roots  than  in  seeds.  Among  the  cereal  products  the  proportion  of 
inorganic  phosphorus  in  the  total  is  much  higher  in  brewer's  grains 
and  malt  sprouts  than  in  others. 

Organic  and  inorganic  phosphorus  in  meat  and  milk  are  some- 
what nearly  equally  divided.  Inorganic  phosphorus  predominates 
in  whey,  while  eggs  are  practically  free  from  inorganic  phosphorus. 

Since  the  whole  matter  of  inorganic  phosphorus  estimation  in 
vegetable  substances  is  in  an  unsettled  state  we  have  made  no  effort 
to  present  all  the  data  available.  Below  are  references  to  a  few 
of  the  investigations  which  report  inorganic  phosphorus  estimations 
in  vegetable  substances : 

Zaleski  (1902,  1907) ;  .Schlagdenhauff en  and  Reeb  (1902) ;  Hart 
and  Andrews  (1903) ;  Schulze  and  Castoro  (1904) ;  Suzuki  and 
Yoshimura  (1907) ;  Suzuki,  Yoshimura  and  Takaishi  (1907) ;  Stut- 
zer  (1908) ;  Heubner  and  Reeb  (1908) ;  Forbes,  Lehmann,  Collison 
and  Whittier  (1910) ;  Forbes,  Whittier  and  Collison  (1910) ;  Hart 
and  Tottingham  (1910) ;  Hartwell  and  Quantz  (1910) ;  Rising 
(1910) ;  Vorbrodt  (1910) ;  Hartwell  and  Hammett  (1911) ;  J.  W. 
Ames  and  Boltz  (1912) ;  Hartwell  (1913). 

PHYTIN  IN  FOODS 

Posternak  (1903a,  1903b,  1905)  found  phytin  in  the  aleurone 
grains  of  red  fir,  hemp  and  sunflower,  in  peas,  lentils,  white  kidney 
beans,  rape,  lupine,  wheat,  corn,  potatoes,  dahlia-bulbs,  carrots  and 
even  onions,  and  says  that  in  grains,  where  there  is  little  mineral 
phosphate,  it  forms  at  least  70-90  percent  of  the  total  phosphorus. 
He  looks  upon  it  as  a  storage  of  reserve  material  for  the  develop- 
ment of  the  embryo.  The  following  table  is  taken  from  the  re- 
ports of  1903.     , 

PHYTIN  AND  LECITHIN  PHOSPHORUS  IN  FOODS  (Posternak,  1903)"*" 


Red    fir     

Hempseed   (cortex  removed) 

Sunflower   seed    (cortex   removed) 

Peas    

Lentils     

White  kidney  beans 


Total  phos- 
phorus,     per- 
cent 


Phosphorus  of 
phytin,   per- 
cent 


0.656 
1.460 
0.830 
0.367 
0.299 
0.512 


Phosphorus  of I  Phosphorus  of 
phytin,  in  per-|    lecithin,  in 
cent    of    total  percent   of  to- 
phosphorus      tal  phosphorus 


0.600 
1.330 
0.723 
0.260 
0.247 
0.418 


91.46 
91.44 
86.26 
70.80 
82.60 
81.60 


1.1 
3.1 

1.8 
6.2 
6.7 
6.0 


82 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


Gilbert  and  Posternak  (1905)  say  that  in  a  mixed  diet 
the  main  part  of  the  phosphorus  is  in  the  form  of  phytin.  It  is 
now  well-known  that  phytin  as  estimated  by  Gilbert  and  Posternak 
includes  compounds  other  than  phytin. 

Suzuki,  Yoshimura  and  Takaishi  (Suzuki  and  Yoshimura,  1907; 
Suzuki,  Yoshimura  and  Takaishi,  1907),  from  their  studies  of  the 
distribution  of  phytin,  report  the  following  determinations  of  the 
forms  of  phosphorus  in  bran,  seeds,  seedlings  and  a  few  other  sub- 
stances. 

PARTITION  OF  THE  PHOSPHORUS  COMPOUNDS  OF  FOODS   (Suzuki, 
Yoshimura  and  Takaishi) — Percent 


Material  examined 


Rice   bran 

Wheat  bran 

Seeds  of  sesame  (Sesamum 

indicutn)   

Seeds  of  castor  bean    (Rtc 

inns  communis) 

Oil  cake,  rape  (Brassica 

napus) 

Barley  bran    {Hordeum 

■vulgar  e) 

Millet  bran  (Panicum 

frumeiitaceum) 

Rice  seed 

seedling's 

Rape  seed 

seedling's 

Barley  seed 

seedlings 

Wheat  seed 

seedlings 

Steamed  bone  meal  (air- 
dry) 

Fresh  bones  of  young    cock 

(1  percent  HC1  used) . . 


Total 

phos- 
phorus 


Phosphorus    in 
lecithin 


In    dry 
sub- 
stance 


2.27 
1.114 

0.772 

0.261 

1.195 

0.541 

0.765 


10.077 
9.186 


In  dry 
sub- 
stance 


0.02 
0.010 

0.030 

0.013 

0.034 

0.010 

0.026 


0.009 


Percent 
of  total 


0.86 
0.81 


3.91 
5.13 


1.85 

3.40 
1.47 
4.10 
6.95 
9.83 
4.14 
8.05 
3.32 
6.97 

0.09 


Phosphorus  sol- 
uble in  0.2  per- 
cent HC1 


Of  that  soluble    in    0.2    percent 
HC1 


In  dry 

sub- 
stance 


Percent 
of  total 


1.92 
0.638 

0.144 

0.110 
0.592 
0.327 
0.363 


10.066 
7.168 


84.48 
57.24 

18.61 

42.29 

49.52 

60.44 

47.45 
44.28 
76.00 
64.51 
78.18 
58.62 
73.34 
56.56 
74.26 

99.89 

78.03 


Inorganic 


In  dry 
sub- 
stance 


0.13 
0.050 


0.089 
trace 


8.919 
6.514 


Percent 
of  total 


5.89 
4.49 


16.45 

trace 

25.22 
trace 
70.02 
trace 
65.98 
trace 
63.27 

88.51 

70.91 


Organic 


In    dry 

sub- 
stance 


1.68 
0.579 

0.125 

0.109 

0.532 

0.238 

0.344 


0.267 
0.167 


Percent 
of  total 


74.17 
52.00 

16.24 

41.61 

44.46 

44.00 

44.97 
41.64 
42.52 
62.11 

2.88 
56.55 

6.89 
55.50 
10.72. 

2.65 

1.82 


The  authors  conclude : 

"1.  The  greater  part  of  the  phosphorus  of  plant  seeds  consists 
of  the  organic  compound  soluble  in  water  and  dilute  mineral  acids 
which  has  been  obtained  by  Schulze,  Palladin,  Winterstein,  Poster- 
nak, Patten  and  others,  and  named  'Anhydro-oxy-methylen-diphos- 
phoric  acid'  or  'Phytin/ 

"From  rice  bran  we  have  isolated  about  8  percent  and  from 
wheat  bran  about  2  percent  phytin. 

"2.  But  in  roots,  bulbs  and  fruits  inorganically  combined 
phosphoric  acid  predominates. 

"3.     The  occurrence  of  phytin  in  bones  is  doubtful. 

"4.  During  the  germination  of  plant  seeds,  either  in  the  light 
or  in  the  dark,  the  inorganically  combined  phosphoric  acid  increases 
notably. 


PHOSPHORUS  METABOLISM  83 

"Also  if  one  grinds  rice  or  wheat  bran  or  different  seeds  and 
lets  them  stand  suspended  in  water  for  some  days,  phosphoric  acid 
forms  in  considerable  quantity  at  the  expense  of  phytin. 

"5.  An  enzyme  was  isolated  from  rice  and  wheat  bran  that 
splits  phytin  into  phosphoric  acid  and  inosite.  It  is  apparently  a 
new  enzyme  and  seems  to  be  widely  distributed  in  the  plant  world." 

Hart  and  Tottingham  (1910)  proved  the  presence  of  such  a 
compound  also  in  the  grains  of  corn,  oats  and  barley,  and  distributed 
throughout  the  entire  seed,  in  these  cereals,  more  than  in  wheat. 
Below  is  their  description  of  the  separation  of  parts  of  the  seeds. 

"In  the  wheat  kernel  phytic  acid  as  a  salt  exists  largely  in  the 
outer  aleurone  layers  and  consequently  is  found  in  very  large  pro- 
portion in  wheat  bran.  In  order  to  determine  whether  similar  dis- 
tribution obtained  in  the  corn  kernel  this  seed  was  mechanically  div- 
ided into  three  parts — the  outer  layer  or  corn  bran  (pericarp) ,  the 
germ,  and  the  starch  and  gluten  cells  (endosperm) ."  The  determin- 
ations below  are  stated  on  an  air-dry  basis. 

Total    P  P    soluble    in     0.2    percent 

Percent  -r,HC1  * 

Percent 

Entire  seed 0.29  0.13 

Corn  bran 0.13  0.00 

Corn  germ 0.38  0.13 

Corn   gluten    0.42  0.15 

"It  is  clear  from  these  data  that  in  the  maize  grain,  phytin  is 
not  localized  in  the  outer  layers In  this  instance  there  ap- 
pears to  be  none  in  the  outer  skin  or  seed  coats,  while  there  is,  on 
the  contrary,  more  or  less  uniform  distribution  throughout  the  en- 
tire seed." 

The  oat  "seed  was  mechanically  separated  into  the  hull  or  bran 
layers  (pericarp)  and  kernel.  The  former  is  fibrous  and  forms  a 
considerable  part  of  the  grain.  The  latter  consists  of  the  aleurone 
layer  and  starch  cells  (endosperm)  and  the  embryo." 

_  ,  ,    _  P  soluble  in  0.2  percent 

Total   P  jjOl 

Percent  Percent 

Entire    seed    0.41  0.18 

Oat  kernel    0.41  0.22 

Oat  hull 0.41  0.09 

"It  is  apparent  from  the  data  that  while  the  seed  coats  carry 
a  total  amount  of  phosphorus  comparable  with  the  other  parts  of 
the  grain,  the  proportion  of  phytin  in  the  outer  layers  is  relatively 
small.  On  the  other  hand  it  constitutes  50  percent  of  the  total 
phosphorus  bearing  bodies  of  the  remaining  parts  of  the  seed." 


84  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Of  barley  "the  separation  of  the  entire  seed  into  the  outer 
fibrous  coats  (pericarp),  here  designated  as  bran,  and  into  the 
aleurone  layer,  starch  cells  and  embryo  (kernel)  was  carried  out 
mechanically." 

Total  P  P  soluble  in  0.2  percent  HCi 

Percent  Percent 

Entire  seed 0.50 0.19 

Bran,  or  hull 0. 22 0.15 

Kernel,  or  germ 0.57 0.17 

"These  results  make  it  manifest  that  phytin  is  not  concentrated 
in  the  outer  seed  coats,  although  it  does  constitute  a  very  large  pro- 
portion of  the  total  phosphorus  existing  there.  Phytin  appears  to 
be  distributed  throughout  the  entire  seed." 

Vorbrodt  (1910),  after  studying  the  distribution  of  the  kinds 
of  phosphorus  compounds  in  maize  seed,  says :  "In  corn  seeds  almost 
nine-tenths  of  the  total  phosphoric  acid  and  almost  all  of  that  of 
the  soluble  organic  compounds  is  in  the  germ  and  the  scutellum. 
In  the  germ  and  scutellum  there  are  large  amounts  of  nucleopro- 
teins,  while  the  rest  of  the  seed  contains  but  little  of  them." 

See  also  Rising  p.  79,  and  Rengniez  p.  78,  Bernardini  p.  79-80. 

DISTRIBUTION  OF  LECITHIN  IN  PLANTS  AND  IN  MISCELLANEOUS 

FOODS 

The  data  of  the  table  below  are  taken  from  Schulze  and  Steiger 
(1889),  Schulze  and  Frankfurt  (1894),  and  Schulze  (1897,  1908b). 
Phosphorus  determinations  were  made  on  the  ether-absolute  alco- 
hol extract  and  these  values  were  interpreted  as  lecithin  (3.84  per- 
cent P).  The  values,  therefore,  must  not  be  taken  as  the  absolute 
content,  but  they  show,  rather,  the  relative  amounts  of  phosphorus 
in  such  a  combination.  Where  determinations  have  been  found  to 
differ  in  the  reports  of  different  years,  that  given  latest  has  been 
taken,  and  the  dates  are  recorded  in  all  cases. 


PHOSPHORUS  METABOLISM 


85 


PHOSPHATID    PHOSPHORUS    IN   VEGETABLE    SUBSTANCES— Percent, 

Dry  Basis 


Substance 
Seeds  of 

Date 

i 

Phosphorus 
in  ether-al- 
cohol   ex- 
tract 

2ther-alco- 
lol    soluble 
phosphorus 
computed 
to  lecithin 

Total  phos- 
phorus 

Phosphatid 

phospho- 
rus in   per- 
cent of 
total 

1908 

1897 
1889 
1889 
1897 
1908 

1897 

1894 
1897 

1908 

1894 
1908 

1897 

a 

1908 

1894 

" 

" 

0.082 
0.084 

0.031 
0.063 

0.049 
0.035 

0.022 

0.017 
0.021 
0.009 
0.011 
0.011 
0.026 
0.026 

0.038 
0.033 
0.059 
0.020 
0.021 
0.019 
0.006 
0.004 
0.009 
0.0x4 
0.001 
0.007 
0.009 
0.025 
0.006 
0.020 
0.028 
0.024 
0.016 
0.039 
0.012 
0.073 

2.14 

2.19 

1.09 

0.81 

1.64 

1.05 

1.27 

0.90 

1.03 

0.43 

0.47 

0.25 

0.57 

0.53 

0.73 

0.85 

0.44 

0.55 

0.25 

0.29 

0.30 

0.67 

0.67 

0.49 

0.27 

0.11 

0.99 

0.86 

1.55 

0.54 

0.56 

0.50 

0.15 

0.10 

0.25 

0.37 

0.04 

0.19 

0.22 

0.69 

0.17 

0.54 

0.77 

0.65 

0.45 

0.86 

0.32 

1.94 

1.53 
1.32 

2.10 
1.14 

1.16 

2.60 

5.5 

3.7 

1.0 

1.0 

Horsechestnut  (Aesculus  hippocastamim) 

3.2 

1.3 

"      II 

"      III 

"      "          "       II 

"     II 

From  the  work  of  Stoklasa  (1896a,  1896b)  we  quote  the  follow- 
ing summary  of  conclusions  as  to  the  distribution  of  lecithins : 

"1.  Roots.  Annual  plants  contain  very  little  lecithin  in  their 
roots,  the  maximum  being  0.3  percent ;  at  completion  of  growth  this 
falls  to  0.1  percent.  Perennials  or  biennials  contain  a  larger  quan- 
tity at  the  close  of  the  season,  this  serving  as  a  reserve  for  use  in 
building  new  cells. 

"2.  Stalk.  Stems  contain  0.3  to  0.4  percent  lecithin :  after  the 
fruit  ripens  this  decreases  rapidly,  being  at  most  0.1  percent  then 
for  annuals. 


8Q 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


"3.  Leaves.  A  considerable  quantity  of  lecithin  is  continually 
present  in  the  leaves ;  it  begins  to  disappear  after  fertilization  and 
during  fruit  formation,  if  at  the  same  time  the  leaves  begin  to  turn 
yellow.  Pure  leaf  substance  is  the  richest  part  of  the  plant  in  leci- 
thin, except  the  anthers  and  pollen  grains  at  the  time  of  blooming. 
Leaves  contain  up  to  40  percent  of  their  total  phosphorus  as  lecithin. 
Since  I  have  observed  that  the  removal  and  destruction  of  lecithin 
and  chlorophyl-coloring  matter  run  parallel,  and  since  I  consider 
it  most  probable  that  this  coloring  matter  contains  a  quantity  of 
phosphorus  corresponding  to  that  of  lecithin,  I  am  of  the  opinion 
that  chlorophyl  itself  is  a  lecithin. 

"4.  Flowers.  The  petals,  it  appears,  contain  the  most  lecithin 
at  the  stage  of  fullest  development  of  the  buds ;  after  fertilization 
the  lecithin  decreases.  The  male  organs,  pollen  filaments,  anthers 
and  pollen  grains,  are  the  parts  richest  in  lecithin.  The  pollen . 
grains,  containing  almost  6  percent  of  lecithin,  are  the  richest  of  all. 
This  corresponds  to  the  fact  that  the  sperm  of  higher  animals  is  es- 
pecially rich  in  lecithin." 

From  the  work  of  Stellwaag  (1890)  we  quote  the  following  es- 
timations of  lecithin  in  the  ether  and  benzene  extracts  of  miscel- 
laneous vegetable  foods. 

LECITHIN  AND  PHOSPHORUS   IN  ETHER   EXTRACT  AND   BENZENE 
EXTRACT   OF    FODDERS— Percent    (Stellwaag,    1890) 


Fat  of 


Hay     

Peas    

Vetch    

Tick   bean    .... 
Lupine    blossom 
Buckwheat 
Soja   bean    .... 

Cabbage    

Poppyseed  cake 
Cottonseed   cake 

Potatoes 

Barley    


Ether  Extract 


Lecithin  Phosphorus 


Traces 

27.37 

22.94 

21.29 

4.59 

1.88 

1.26 

6.99 

13.27 

4.35 

3.07 

4.25 


Traces 
1.049 
0.881 
0.818 
0.172 
0.072 
0.066 
0.268 
0.40 
0.166 
0.117 
0.163 


Benzene   Extract 


Lecithin 


6.95 
7.65 
4.11 

2.53 
1.57 
3.27 
6.24 
1.52 

2.37 


Phosphorus 


0.247 

0.26 

0.14 

0.083 
0.063 
0.123 
0.222 
0.058 

0.09i 


Von  Bitto  (1894)  prefers  a  methyl  alcohol  extraction  (20  ex- 
tractions) to  the  method  of  Schulze  and  Steiger,  for  phosphatid  de- 
termination, and  reports  the  following  amounts  found  in  that  way : 


PHOSPHORUS  METABOLISM 


87 


LECITHIN  PHQSPHORUS  AND  LECITHIN  IN  VARIOUS  SEEDS— Percent 

Dry  Basis 


P 

Lecithin 

0.0687 
0.0621 
0.0746 
0.0750 
0.0222 
0.0256 
0.0227 
0.0185 

1.788 

1.618 

Lupinus   luteus    (yellow   lupine)     .  .  . 

1.933 
1.955 

0.578 

0.667 

0.592 

0.482 

Schulze  (1895)  supports  his  method  in  comparison  with  that 
of  von  Bitto. 

Th.  Dietrich  (1902)  reports  1.48  to  1.89  percent  of  lecithin  in 
the  dry  substance  of  brewer's  grains. 

Stoklasa  (1895)  found  in  sugar  beet  leaves  1.12  percent,  and  in 
the  root  0.43  percent  lecithin,  on  the  dry  basis. 

W.  Koch  (1905a)  reported  lecithin  estimations  on  a  number  of 
common  foods,  as  below: 


LECITHIN  IN  SOME  COMMON  FOODS  (W.  Koch,  1905) 


1  lb.  calf  brain  .  . 
1  lb.  shad  roe  .... 

12   eggs    

1  lb.  calf  liver  . . . 
1  lb.  sweetbreads  . 
1  lb.  lamb  fries  .  . 
1  lb.  meat  (beef)  . 
1  lb.  peas  or  beans 

1  lb.  salmon    

1  lb.  bread   

1  lb.  vegetables  .  . 
1  pint  human  milk 
1  pint  bovine  milk 
1  lb.  mushrooms   .  . 


Gm.  lecithin 


20-25 

18-20 

12-16 

12-15 

12-15 

10-12 

5-7 

5-7 

5-6 

0.5-1 

0.3-0.5 

0.4 

0.3 

0.2 


Heubner  and  Reeb  (1908)  made  determinations  of  the  amount 
of  phosphorus  in  the  forms  of  its  different  compounds  in  a  number 
of  common  foods,  and  their  table  is  given  below.  Heubner  (1911) 
says  later  that  the  method  used  here  is  inexact  for  the  estimation 
of  organic  phosphorus,  for  instance,  phytic  acid. 


88 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


Q 

O 
O 

ft 


<5 

H 

Woo 

w  OS 

5- 

S  s 

H  * 
P  n 

as 

H  « 
COM 
i— i " 

ft 


I— I 

o 
w 

o 

M 


ri 

,  d 

»  o  ° 

COt-OsaoONWCOlNCO 

u-3o 

Nino^cccoooHO 

2  ft  M 

oooooooooo 

oooooooooo 

ra  £,D 

<J  o!  p, 

c3        CD  P 

^^ 

OCOCOO     •tomi-HCD 

rHOOO     .t)IHOHO 

Ph 

OOOO     -coOOOO 

dodo      OOOOO 

ea 

•             — •' — 

-5    1 

03 

.3     a> 
5    1 

-HMMlO^t-t-Nt-tSI 

HMOMOTfCilHINH 

O         ft 

rHOOOOcNOOOO 

,£] 

VI          VI 

odddddodoo 

VI 

a 

u 

3  -ft 

a 

o    3 

ft       °3 

OiCOOt--d(OTilCDOMO 

MOOOONOOOO 

. 

ooooooooco 

H 

2     ft 

oooooooooo 

CTtOiO00t-<N-^0000<O     , 

"3 

050tH050^LOVOCNmcN 

If 

iHrHOOOrfOOOO 

En 

OOOOOrHOOOO 

.2    '   .2 

"<u    §   'a 

OOt-t-Ot-*NOrf 

1 

2  "S    o      ■ 

ri^OOHOOOHO 

^        ft      fH 

S     n-t       ft 
*0      o 

OOOOOOOOOO 

co     fl    A- 

<j      °3      ft 

li 

^^^ 

5  o    t, 

^   =>    "5 

r^iOCNO    •lOT-HiOOlt^ 

OOOO     .(OHOOO 

dddd   'ddddd 

o      » 

v^^- 

05      « 

P4 

-91 

— ' 

O                 4^ 
r3                ft 

w 

THiOtMLOOmOOOtM^II 

o 

rfNOOONNOlMH 

rt 

O          vx 

OOOOOOOOOO 

03 

02             O 

A 

PS 

%     ft 

Ed 

*>> 

§     3 

in 

mmoHofico^t-© 

H 

i-IOOOOOOOOO 

"O 

ft          °3 

OOOOOOOOOO 

CD 

VI             Oi 

rf 

rt 

<] 

M 

T3 

PS  O  iH  CO  rH  Cl  00  O     •     • 

<o  ri 

t- 00  iH  rH  rH  CM  CO  rH      •      • 

C3  _0 

odddoHod 

F4 

03 

T3 

o 

t      CD 

En 

£« 

■"t^CQ^Ht-OOOOH 

Si 

t-OOWrHr-I^TlliH^CO 

OOOOOiHOOOO 

5  ® 

>d 

•2 

0        © 
03 

OCOCDr-tOOOOCOCOCOCO 

doicsidc^odcOTiicsiod 

CNrHrHt-OOairHiHiH 

0 

bO 

M 

<D 

03 

•a 

VI 

CD    CD 

CD 

■3 

"a 

M  03   03 
iS  A  A 

3 

f&A 
S;  03  03 

tiao    "    ' 

to^i  o  o 

"       ft 
(DtatBfcj           .JT^flCD 

c/ 

z 

n 

> 

a1 

E- 

r 

bi 

tz: 

CDS 

'  £  A 

PHOSPHORUS  METABOLISM 


89 


Vageler  (1909)  finds^  as  do  others,  that  seeds  contain  but  a 
small  amount  of  inorganic  phosphorus ;  also  that  the  largest  amount 
of  phosphatid  is  to  be  found  in  a  plant  at  the  time  of  its  fullest  veg- 
etation and  while  the  seeds  are  forming;  when  autumn  comes  and 
the  seeds  have  ripened  and  leaves  are  withering,  the  easily  soluble 
phosphorus  is  low;  also  that  the  phosphorus  content  of  a  normal, 
healthy  plant  is  greater  than  that  of  an  ill-nourished  plant.  Vageler 
also  questions  whether  it  may  not  be  that  the  low  values  that  ob- 
servers have  found  for  the  phosphorus  content  of  the  ether-alcohol 
extract  of  dried  vegetable  products  is  due  to  a  partial  breaking  up 
of  phosphatid  during  the  drying  process. 

ANALYSES  OF  FRESH  AND  DRIED  VEGETABLE  TISSUE  FOR  WATER 
AND  PHOSPHATID  PHOSPHORUS    (Vageler,   1909)— Percent 


Fresh  material 

Dried  rr 

aterial 

Substance  examined 

Water 

Phosphatid 
phosphorus 

Water 

Phosphatid 
phosphorus 

94.93 

89.74 

90.165 

85.55 

14.74 

17.26 

14.94 

0.1487 
0.0376 
0.2934 
0.0780 
0.0169 
0.0496 
0.0203 

10.37 
9.596 
7.218 
4.898 

12.98 
8.03 

11.66 

0.0731 

0.0353 

0.0578 

0.584 

Hay 

0.0091 

Lupine  seeds. . .  > 

0.0398 
0.0189 

Winterstein  and  Smolenski  (1909)  find  that  the  phosphatid  ob- 
tained from  wheat  flour  is  a  mixture  of  at  least  three  phosphatids 
with  cholesterin  and  other  substances;  and  K.  Smolenski  (1909) 
isolated  two  phosphatids  from  wheat  germ. 

LeClerc  and  Wahl  (1909)  report  finding  considerable  loss  of 
phosphoric  acid  and  of  other  ash  constituents  in  the  change  from 
barley  to  malt.  At  the  same  time  there  was  an  increase  of  alco- 
hol-ether soluble  phosphorus,  and  probably  a  part  of  the  other  forms 
of  organic  phosphorus  are  transformed  to  phosphatids,  while  con- 
siderable, as  others  have  found,  becomes  inorganic.  Of  the  130 
barleys  analyzed  the  general  average  of  total  P,05  is  1.04  percent, 
and  of  lecithin  0.52  percent  in  the  water-free  .substance; 
and  of' 43  malts  the  general  averages  are  1.03  percent  total 
phosphoric  acid  and  0.75  percent  lecithin.  The  average  percent  of 
alterations  during  malting  are  computed  to  a  loss  of  12.7  percent 
of  the  total  phosphorus  and  a  gain  of  44.3  percent  of  the  lecithin. 

Fraps  and  Rather  (1912)  have  investigated  the  composition  of 
the  ether  extract  of  several  kinds  of  hay,  straw  and  other  roughage. 
The  phosphorus  content  of  the  ether  extract  may  be  considered 
roughly  to  indicate  the  phosphatid  content  of  the  products.    From 


90  OHIO  EXPEEIMENT  STATION:  TECHNICAL  BUL.  5 

these  figures  it  would  appear  that  the  phosphatid  content  of  these 
species  of  roughage  is  low,  as  indeed  all  roughage  is,  and  also  that 
peanut  hay  is  distinctly  richer  in  ether-soluble  phosphorus  than  the 
other  kinds  of  hay  studied.      Below  are  figures  from  this  study. 

NITROGEN  AND  PHOSPHORIC  ACID  OF  THE  ETHER   EXTRACT  OF 
HAYS  AND  FODDERS  (Fraps  and  Rather,  1912) 


Feed 


Alfalfa   hay    

Bermuda    hay    

Buffalo  grass  hay 

Burr  clover    

Corn    shucks    

Cowpea  hay 

Guam    grass     

Johnson   grass   hay    .  .  . 
Johnson   grass   hay    .  .  . 

Kafir   fodder    

Millet    

Oat  hay    

Para  grass  hay   

Peanut   hay    

Rice  straw,  Japan    .  .  .  . 
Rice   straw,    Honduras 

Sorghum      

Vetch  hay    


Total    ether      extract 
Percent 


1.26 
1.55 
1.28 
2.72 
0.61 
3.15 
1.78 
1.29 
1.38 
1.99 
1.53 
2.12 
0.86 
8.17 
1.47 
1.24 
1.47 
1.59 


Percent  of  extract 


Nitrogen 


0.23 
0.38 
0.33 
0.44 
0.20 

0.56 
0.26 
0.34 
0.39 
0.19 
0.33 
0.23 
0.07 
0.26 
0.18 
0.23 
0.42 


Phosphoric   acid 


0.06 
0.08 
0.06 
0.10 
0.19 
0.13 
0.14 
0.09 
0.15 
0.10 
0.09 
0.05 
0.25 
0.18 
0.02 
0.06 
0.09 
0.06 


See  also  Posternak,  p.  81,  Rising,  p.  79,  Suzuki,  Yoshimura  and  Takaishi, 
p.  82,  Vorbrodt,  p.  79,  and  Bernardini,  p.  79-80. 

DISTRIBUTION  OF  NUCLEIN  PHOSPHORUS  IN  FOODS 

There  are  no  well-established  methods  for  the  routine 
estimation  of  nuclein  phosphorus  in  miscellaneous  foods.  Nuclein 
phosphorus  has  been  estimated,  however,  by  various  more  or  less 
satisfactory  methods,  and  for  the  results  of  some  of  these  determin- 
ations we  would  cite  the  work  of  Heubner  and  Reeb  (1908),  Jebbink 
(1910)  (60  articles  of  human  diet)  and  Jordan,  Hart  and  Patten 
(1906). 

In  Klinkenberg's  (1882)  study  of  nucleins  he  gives  us  the  fol- 
lowing as  the  content  of  nuclein  phosphorus  in  seed  cakes  com- 
pared with  meat  residues  and  with  yeast. 

NUCLEIN    PHOSPHORUS    OF    SEED    CAKES    AND    MEAT    RESIDUES 

Klinkenberg  (1882) 


Poppy  seed   cake    

Peanut   cake    

Rape  seed  cake  

Cotton  seed  cake    

Palm  cake   

Residue  from  meat  extract  preparation  I.  . 
Residue  from  meat  extract  preparation  II. 
Yeast     


Nuclein   P 
Percent 


0.0707 
0.0360 
0.0676 
0.0630 
0.0344 
0.0307 
0.0530 
0.198 


PHOSPHORUS  METABOLISM 


91 


About  3.5' percent  of  wheat  germ  is  tritico-nucleic  acid,  which, 
was  first  studied  by  T.  B.  Osborne  and  Campbell  (1900a),  and 
later  by  T.  B.  Osborne  and  Karris  (1902)  and  Levene 
and  LaForge  (1910).  This  acid  is  different  in  its  makeup  from 
the  animal  nucleic  acids.  Osborne  and  Campbell  isolated  the  acid 
itself  and  also  obtained  combinations  of  it  with  leucosin,  a  globulin 
and  two  proteoses,  as  well  as  showing  that  some  of  it  was  left  in 
combination  with  other  insoluble  substances.  With  regard  to  the 
combinations  in  the  wheat  embryo,  however,  it  is  said  finally :  "That 
the  wheat  embryo  in  fact  contained  the  same  nucleic  acid  compounds 
as  we  have  obtained  from  the  extracts  is  highly  improbable.  All 
that  we  can  conclude  is  that  the  embryo  contains  the  different  pro- 
tein substances  described,  together  with  nucleic  acid,  and  that 
these  may  unite  to  form  a  number  of  different  compounds  according 
to  the  conditions  which  prevail  at  any  given  time." 

Funatsu  (1907)  also  estimated  nuclein  phosphorus  in  oil  cakes. 
The  following  data  are  submitted : 

DETERMINATIONS    OF    THE    PHOSPHORUS    PARTITION    IN    PRESS 

CAKES  (Funatsu,  1907) 


Total 
P2O5 

Lecithin  P2Os 

Nuclein  P2O5 

P0O5     soluble     in. 
dilute  HC1 

Percent 

Percent 
of  total 

Percent 

Percent 
of   total 

Percent 

Percent 
of   total 

1.38 
2.25 
2.82 

0.17 
0.12 

0.20 

12.4 

5.0 
7.0 

0.23 
0.30 
0.26 

16.5 

13.2 

9.0 

0.98 
1.80 
2.37 

71.0 

81.7 

84.0 

See  also  Rising  p.  79,  Rengniez  p.  78  and  Bernardini  p.  79-80. 

PYROPHOSPHORIC  ACID  IN  VEGETABLE  SUBSTANCES 

The  question  of  the  presence  of  pyrophosphoric  and  metaphos- 
phoric  acids  in  cottonseed  meal,  first  raised  by  Hardin  (1892),  has 
called  forth  investigation,  especially  because  Crawford  (1910) 
thought  that  he  had  evidence  that  the  poisonous  effects  sometimes 
observed  as  a  result  of  feeding  cottonseed  meal  to  cattle  and  hogs 
is  due  to  the  presence  in  the  meal  of  a  salt,  either  organic  or  inor- 
ganic, of  pyrophosphoric  acid.  The  evidence  of  the  presence  of 
these  acids  is  in  qualitative  reactions  in  extracts  of  cottonseed  meal, 
which  resemble  those  of  the  acids  in  question,  and  in  toxic  effects 
of  the  meal,  or  extracts  from  it,  which  are  similar  to  those  of  the 
acids.  More  recently,  however,  both  Rather  (1912)  and  Anderson 
(1912c)  have  independently  shown  that  all  of  the  reactions  in  ques- 
tion are  given  by  organic  phosphorus  compounds,  salts  of  which 


92  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

they  have  isolated  from  cottonseed  meal. '  Both  find  the  acids  of 
the  salts  isolated  to  be  inosite  phosphoric  acid  (Rather  1913b,  An- 
derson 1914a).  It  is  not  markedly  toxic  in  small  doses.Withers  and 
Ray  (1913)  have  shown  that  the  only  fraction  of  cottonseed  meal 
which  is  toxic  to  rabbits  in  the  amounts  used  in  feeding  is  that 
which  is  not  soluble  in  water  or  dissolved  by  digestion  with  pepsin 
and  pancreatin  (one  day  each),  and  that  if  this  fraction  contains 
any  pyrophosphoric  acid  it  is  a  non-toxic  amount.  It  is  to  be  con- 
cluded that  probably  there  is  no  pyrophosphoric  acid  in  this  meal; 
also  that  the  principal  phosphorus  compound  present  is  organic,  and 
of  the  nature  of  phytin. 

PROPRIETARY  PREPARATIONS 

Phosphorus  compounds  as  constituents  of  medicinal  prepara- 
tions have  occupied  a  prominence  out  of  proportion  to  their  useful- 
ness in  this  relation.  While  they  are  of  undoubted  value  as  simple 
nutrients,  the  idea  of  their  possession  of  a  stimulating  function  by 
reason  of  direct  contribution  to  the  nervous  tissues,  or  by  reason  of 
other  specific  effects,  has  been  very  greatly  overworked.  It  is  true, 
however,  that  there  is  at  hand  evidence  that  certain  of  the  organic 
compounds  of  phosphorus,  lecithin  especially,  may,  in  certain  path- 
ological conditions,  have  a  curative  value;  but  even  in  these  cases 
there  is  need  of  further  experimental  work  to  differentiate  between 
the  effects  of  these  compounds  in  the  uncombined  state,  and  the  ef- 
fects of  the  related  compounds  as  they  exist  in  natural  foods.  We 
put  no  emphasis,  therefore,  on  the  usefulness  of  proprietary  medic- 
inal preparations  as  affected  by  their  phosphorus  compounds,  but 
merely  enumerate  those  which  have  come  to  our  attention,  with 
brief  notes  as  to  their  general  character. 

Springer  (1894,  1902)  prepared  a  cereal  decoction  which  he 
recommends  to  be  used  with  the  diet,  especially  in  cases  of  retarded 
growth.  It  is  prepared  by  boiling  a  mixture  of  wheat,  barley,  oats, 
rye,  maize  and  bran  in  water  and  straining  out  the  residues. 

Mouneyrat  (1902a,  1902b)  prepared  what  he  called  "Histo- 
genol,"  a  combination  of  methyl  arsenate  of  sodium  and  the  nucleic 
acid  from  herring  milt. 

Among  casein  preparations  are  "Plasmon"  (E.  Bloch,  1900 ;  Poda 
and  Prausnitz,  1900;  Micko,  1900)  and  "Nutrose,"  impure  sodium 
salts  of  casein,  "Eucasein,"  an  ammonium  salt  described  by  Salkow- 
ski  (1896b),  "Sanose,"  80  percent  casein  and  20  percent  albumose 
(Schreiber  and  Waldvogel,  1897),  and  "Sanatogen,"  sodium-casein 
glycerophosphate  (Snowman,  1905;  Gumpert,  1905). 


PHOSPHORUS  METABOLISM 


93 


Special  glycerophosphate  and  lecithin  preparations  are  dis- 
cussed by  Frey  (1906),  especially  Baraba's  "Nervinol." 

Laves  (1900)  describes  "Roborat,"  a  cereal  preparation  said  to 
contain  an  abundance  of  lecithin,  and  Heim  (1904)  describes  "Bio- 
son"  as  a  protein-iron-lecithin  compound  containing-  1.27  percent 
lecithin. 

Schroder  (1905, 1906)  describes  "Bioplastin,"  a  lecithin  prepar- 
ation, "Histogenol,"  and  "Caudol,"  a  dry  malt  extract  containing 
phosphates. 

"Fersan,"  prepared  from  the  blood  of  cattle,  contains  iron  and 
phosphorus  organically  combined  (Kornauth  and  Czadek,  1900 ;  Kor- 
nauth,  1901). 

"Protylin"  is  a  synthetic  paranuclein  containing  about  2.7  per- 
cent phosphorus  (Kornfeld,  1904;  Laguesse,  1905;  Fjodoroff,  1907). 

EFFECTS  OF  WATER  ON  THE  COMPOSITION  OF  FORAGE  PLANTS 

Kellner,  Kohler  and  Barnstein  (1894,  1895)  give  data  which 
show  that  both  hay  and  straw  were  low  in  the  phosphoric  acid  con- 
tent of  the  ash  after  the  dry  summer  of  1893;  and  the  following 
figures  from  von  Seelhorst, -Georgs  and  Fahrenholtz  (1900)  show  a 
like  effect  in  clover  and  grass,  with  moisture  controlled  by  irrigation. 


PHOSPHORIC  ACID  CONTENT  OF  CLOVER  AND  HAY  AS  AFFECTED  BY 

THE  AMOUNT  OF  MOISTURE  SUPPLIED  (Von  Seelhorst  et  al.,  1900) 

Percent  Phosphoric  Acid 


Water 

Plot  manured 

Plot  not  manured 

0.456 
0.536 
0.540 
0.725 
0.597 
0.666 

0.428 
0.553 
0.523 
0.660 
0.644 
0.650 

Clover 

Much    

91 

Little   

Meadow    fescue 

»» 

Forbes,  Whittier  and  Collison  (1910)  showed  in  experiments 
with  oats,  grown  with  various  amounts  of  water,  that  there  is  a  re- 
lation between  the  phosphorus  content  of  the  plant  and  the  water 
available  during  growth.      The  numerical  data  are  below : 


94 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


ANALYSES  OF  OAT  PLANTS  GROWN  WITH  DIFFERENT  AMOUNTS  OF 
WATER— Dry  Matter  Basis— Percent 


Lot 


Total     water     ap- 
plied to  each  pot 

C.    C. 


Moisture   in 
plant 

Percent 


Ash    in    dry    sub- 
stance 

Percent 


Phosphorus   in 
dry    substance 

Percent 


Calcium     in     dry 
substance 

Percent 


Planted  Oct.  7,  1908;  cut  Nov.  20,  19C 


1 

1675 

86.11 

16.77 

.884 

.526 

2 

2420 

88.51 

15.38 

.898 

.451 

3 

3165 

88.20 

15.12 

.966 

.461 

4 

3910 

87.77 

14.72 

.979 

.515 

5 

4655 

88.06 

13.96 

.892 

.453 

Planted  Dec.  13,  1909;  cut  Feb.  26,  1910 


6 

1800 

83.78 

15.17 

.647 

.440 

7 

3100 

84.57 

15.17 

.666 

.409 

8 

4400 

86.79 

15.65 

.805 

.498 

9 

5700 

87.28 

16.02 

.787 

.463 

10 

7000 

87.75 

16.20 

.855 

.463 

Planted  Dec.  13,  1909;  cut  Mar.  7,  1910 


11 

2400 

79.68 

15.37 

.684 

.461 

12 

4000 

82.44 

16.19 

.768 

.504 

13 

5600 

85.94 

16.20 

.801 

.495 

14 

7200 

85.18 

16.18 

.818 

.488 

15 

8800 

84.76 

15.98 

.809 

.480 

Planted    Apr.    4,    1910;    cut    May    11,    1910 

16 

900 

88.97 

18.47 

1.12 

.542 

17 

1425 

89.59 

18.68 

1.16 

.539 

18 

2000 

89.95 

18.54 

1.20 

.554 

19 

2625 

90.50 

18.20 

1.22 

.493 

20 

3300 

90.73 

18.05 

1.27 

.551 

A  similar  relationship  between  water  and  phosphorus  was 
shown  by  these  same  authors  in  forage  plants  grown  in  arid  regions 
with  and  without  irrigation.      The  data  are  below : 

EFFECTS  OF  IRRIGATION  ON  MINERAL  CONSTITUENTS  OF  GRASSES 
Dry  Matter  Basis — Percent 


No. 

Ash 

Nitrogen 

Calcium 

Potas- 
sium 

Phos- 
phorus 

Source   of   sample 

1 

8.62 

2.32 

1.67 

1.80 

.175 

Alfalfa;  Fallon,  Nev.;  not  irrigated  for 
five  years;  water  table  8  ft.  below  sur- 
face of  field. 

2 

10.34 

2.50 

1.27 

2.38 

.220 

Alfalfa;  Fallon,  Nev.;  irrigated  frequently. 

3 

16.38 

.845 

.342 

.99 

.099 

Blue-joint  (Elymus)  ;  Fallon,  Nev.;  sample 
from  dry  soil. 

4 

9.47 

766 

.250 

1.06 

.133 

Blue-joint  (EZy?nus);  Fallon,  Nev.; 
sample  from  land  continuously  wet  by 
seepage  from  irrigation   ditch. 

5 

9.78 

.895 

.410 

.92 

092 

Indian  bunch-grass,  (JErt'ocoma  cuspidatd) 
Fallon,  Nev.;  sample  from  dry  soil; 
annual  rain-fall  less   than  three  inches. 

6 

16.10 

.734 

.462 

.67 

.103 

Indian  bunch-grass,  (Eri'ocoma  cuspidata) 
Fallon,  Nev. ;  sample  from  land  con- 
stantly wet  by  seepage  from  irrigation 
ditch. 

7 

12.11 

1.89 

1.00 

2.14 

.142 

Bermuda  grass.  Tuma,  Ariz.  Not  irrigated. 

8 

11.46 

1.39 

.709 

1.55 

.243 

Bermuda  grass.  Yuma,  Ariz.  Irrigated 
frequently. 

PHOSPHOEUS  METABOLISM 


95 


EFFECTS  OF  FERTILIZERS  ON  THE  COMPOSITION  OF  FOODS 

Since  some  of  the  phosphorus  compounds  of  plants  are  readily 
soluble  in  the  water  which  falls  upon  them  as  rain,  and  since  we  do 
not  know  that  the  salts  absorbed  by  the  fertilized  plants  are  the 
same  and  therefore  have  the  same  solubility  as  those  absorbed  by 
the  unfertilized  plants,  we  have  no  assurance  that  samples  collected 
from  plants  which  have  been  grown  out-of-doors  fairly  represent 
the  effects  of  the  fertilizers  on  the  phosphorus  compounds  of  the 
plants.  In  the  interpretation  of  numerical  data  on  this  matter, 
therefore,  it  would  seem  wise  to  make  liberal  allowance  for  experi- 
mental error.  It  seems  altogether  probable  that  the  apparent  in- 
consistencies in  results  which  have  been  obtained  are  due  in  part  to 
the  factors  above  suggested.  From  the  mass  of  material  available 
we  select  but  a  small  portion. 

From  the  work  of  Chavan  (1908)  we  quote  the  following  fig- 
ures showing  direct  effects  of  fertilizers  on  the  composition  of  grass. 


INFLUENCE  OF  PHOSPHORUS  AND  POTASSIUM  IN  FERTILIZERS  ON 

THE  COMPOSITION  OF  GRASSES  (Chavan,  1908) 

Percent — Dry  Basis 


Fertilizer 

applied 

Protein 

P2O5 

K20 

CaO 

Ash 

P    

10.29 

8.90 

11.06 

10.25 

0.25 
0.56 
0.24 
0.57 

1.41 

1.80 
2.56 
2.61 

1.52 
1.20 
1.10 
0.89 

6.47 

6.05 

K     

6.82 

P,   K    

7.21 

Parrozzani  (1908)  states  that  increasing  the  inorganic  phos- 
phorus available  to  maize  increases  the  lecithin  and  phytin  phos- 
phorus, but  does  not  affect  nuclein  phosphorus  nor  total  nitrogen, 
though  protein  nitrogen  increases  slightly  and  amid-nitrogen  de- 
creases correspondingly. 

Forbes,  Whittier  and  Collison  (1910)  made  analyses  of  24 
samples  of  Kentucky  blue-grass  taken  in  1910  from  a  variety  of 
types  of  soil  in  Ohio  and  Kentucky,  the  conditions  of  taking  the 
samples  and  the  age  of  the  plants  being  as  nearly  alike  as  possible. 
The  range  of  phosphorus  content  is  from  0.164  to  0.403  percent  P 
in  the  dry  matter.  Samples  taken  the  two  previous  years  were  all 
within  the  same  range.  The  inorganic  phosphorus  in  17  of  the 
samples  ranged  from  0.064  to  0.  267  percent,  and  the  organic,  from 
0.064  to  0.154  percent  of  the  dry  matter.  Blue-grass  from  experi- 
ment plots  differently  fertilized  gave  results  as  in  the  table  below. 


96 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


Potassium  chloride  appears  to  have  increased  the  organic  phospho- 
rus of  the  grass.  The  effects  of  the  fertilizers  are  generally  direct 
and  consistent. 

ANALYSES  OF  BLUE-GRASS  DIFFERENTLY  FERTILIZED 
Forbes,  et    al.  (1910)  Percent  of  Dry  Matter 


No. 

Ash 

Nitro- 
gen 

Cal- 
cium 

Potas- 
sium 

Phos- 
phorus 

Inor- 
ganic 
phos- 
phorus 

Organ- 
ic 
phos- 
phorus 

Source    of    Sample 

1 

2 
3 

4 

'    5 

5.48 
5.08 
5.47 
5.01 

6.16 

1.49 
1.52 
1.50 
1.46 

1.48 

.230 
.247 
.222 
.311 

.250 

1.83 
1.58 
1.86 
1.63 

1.82 

.277 
.235 
.265 
.236 

.264 

.176 
.135 
1.38 
.130 

.161 

.101 
.100 
127 
.106 

.103 

Wooster,  O.      Station  lawn.     Fertilized 
with   sodium   phosphate. 

Wooster,    O.        Station  lawn. 

No  fertilizer. 

Wooster,    O.        Station   lawn. 
Fertilized  with  potassium  chloride. 

Wooster,  O.     Station  lawn.     Fertilized 
with  lime   (calcium  oxide). 

Fertilized  with  sodium  phosphate,  'po- 
tassium  chloride   and  lime    (calcium 
oxide). 

Lewoniewska  (1911)  reports  observations  on  the  phosphorus 
content  of  oats  (grain)  as  affected  by  the  soil.  Protein  and  lecithin 
phosphorus  varied  little,  but  both  phytin  and  phosphates  varied 
much.  With  variation  in  the  phosphorus  of  the  soil  the  relation 
of  total  phosphoric  acid  to  nitrogen  varied  between  100:50  and 
100 :32,  while  the  relation  of  phosphorus  compounds  soluble  in  1  per- 
cent acetic  acid  varied  between  100:20  and  100:6. 

From  the  reports  of  Ames  and  his  associates  (J.  W.  Ames, 
1910;  J.  W.  Ames,  Boltz  and  Stenius,  1912)  with  regard  to  the 
effects  of  fertilizers  on  the  phosphorus  content  of  wheat  we  quote 
several  tables : 

EFFECT  OF  SEASON  AND  PLANT  FOOD  SUPPLIES  ON  THE  PHOS- 
PHORUS AND  NITROGEN  CONTENT 


Phosphorus  content  of  wheat  grain  and  straw— Percent 


•  1904 

1907 

1908 

Maximum 

variation  due 

to  season 

Grain 

Straw 

Grain 

Straw 

Grain 

Straw 

Grain 

Straw 

.395 
.366 
.029 

.072 

.105 

-.033 

.3839 
.3053 
.0786 

.0563 
.0548 
.0015 

.3545 
.3234 
.0311 

.0364 

.0469 

-.0105 

.0405 
.0607 

.0356 

.0581 

PHOSFHORUS  METABOLISM 


97 


AVERAGE  PERCENTAGE  OF  PHOSPHORUS  IN  THE  WHEAT  GRAIN  AT 

WOOSTER 


Treatment 

Phosphorus  in  grain 

Percent 
.3143 

.3080 

.3690 

.3332 

.3615 

AVERAGE  PERCENTAGE  OF  PHOSPHORUS  IN  THE  WHEAT  GRAIN  AT 

STRONGSVILLE 


Tear  and  cross   dressing 

Plot  treatment 

1908 

1909 

Floats 

Lime 

Floats 

Lime 

.4059 
.4168 
.4449 
.4572 
.4287 

.3270 
.2837 
.4122 
.3643 
.4173 

.4646 
.4182 
.4689 
.4671 
.4736 

.2947 

.2775 

Phosphorus,  without  nitrogen  or  potassium .  . 
Phosphorus,  potassium  and  nitrogen 

.4116 
.3845 
.4522 

From  the  extended  studies  from  which  the  above  figures  are 
taken  Ames  concludes  in  part  as  follows : 

"The  composition  of  the  wheat  crop  grown  on  the  unfertilized 
plots  of  two  soils,  containing  different  amounts  of  phosphorus,  po- 
tassium and  nitrogen,  is  in  accordance  with  the  composition  of  these 
soils. 

"The  proportion  of  phosphorus,  potassium  and  nitrogen  in  the 
wheat  plant  is  increased  by  the  addition  of  these  elements  to  the 
soil. 

"Although  the  extent  of  variation  due  to  seasonal  conditions  is 
greater  than  that  produced  by  changes  in  the  composition  of  the 
soil,  the  variations  due  to  soil  treatment  are  relatively  the  same  for 
the  different  seasons. 

"Phosphorus  applied  to  soil,  showing  a  deficiency  of  this  ele- 
ment as  measured  by  crop  yields,  increases  the  amount  of  phos- 
phorus in  the  grain.  Associated  with  this  increased  accumulation 
of  phosphorus  there  is  an  increased  quantity  of  potassium  and  a 
decreased  amount  of  nitrogen. 


98  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

"The  addition  of  lime  to  the  soil  increases  the  amount  of  phos- 
phorus assimilated  by  the  wheat  plant.  With  this  increase  in  the 
phosphorus  content  there  are  the  same  variations  in  the  nitrogen 
and  potassium  as  are  produced  by  the  addition  of  phosphorus. 

"The  composition  of  the  wheat  crop  from  plots  on  the  same  soil 
treated  with  five  different  carriers  of  phosphorus,  namely:  acid 
phosphate,  bone  meal,  dissolved  bone  black,  basic  slag,  and  barn- 
yard manure,  shows  that  the  phosphorus  content  of  the  wheat  plant 
has  been  increased  to  the  greatest  extent  by  manure. 

"The  application  of  untreated  rock  phosphate  to  a  soil  well 
supplied  with  nitrogen  and  potassium,  increases  the  phosphorus  con- 
tent of  the  wheat  plant  to  a  marked  extent. 

"The  percentage  of  nitrogen  in  the  wheat  plant  varies  with  the 
supply  at  its  disposal,  and  is  also  influenced  to  a  considerable  extent 
by  the  supply  of  phosphorus." 

"A  comparison  of  the  composition  of  the  wheat  plant  grown  on 
the  same  soil,  under  different  conditions  of  fertilization,  gives  a 
better  indication  of  the  available  supply  of  nitrogen,  phosphorus  and 
potassium  in  the  soil  than  can  be  obtained  from  the  analysis  of  the 
soil  itself." 

From  a  study  by  J.  W.  Ames  and  Boltz  (1912)  of  the  effects  of 
fertilizers  on  the  composition  of  alfalfa  we  quote  the  table  on  the  fol- 
lowing page.  From  the  study  from  which  these  figures  were  taken, 
Ames  and  Boltz  conclude  in  part  as  follows : 

"The  phosphorus  supply  of  the  soil,  as  increased  by  the  addition 
of  acid  phosphate,  is  reflected  by  the  phosphorus  content  of  the  crop, 
which  follows  the  same  order  as  the  yields  obtained. 

"When  the  fertilizer  used  contained  both  phosphorus  and  nitro- 
gen, the  increase  in  the  amount  of  phosphorus  over  that  found  in 
the  crop  from  unfertilized  soil  is  not  as  great  as  where  phosphorus 
without  nitrogen  was  applied. 

"From  43  to  63  percent  of  the  total  phosphorus  present  in  the 
alfalfa  plant  is  combined  as  organic  phosphorus.  There  is  in  each 
instance,  for  every  condition  of  fertilization,  a  greater  proportion 
of  the  total  phosphorus  present  as  organic  phosphorus  in  the  second 
cutting,  while  the  amount  of  total  phosphorus  is  always  larger  in  the 
first  cutting.  The  water-soluble  phosphorus,  which  includes  prac- 
tically all  the  inorganic  phosphorus  and  a  considerable  portion  of 
that  in  organic  combination,  stands  in  the  same  order  as  the  total 
phosphorus  and  is  present  in  greater  amounts  in  the  crop  of  the  first 
cutting.  The  quantities  of  pepsin-insoluble  phosphorus  which  is 
combined  with  nitrogen  as  a  highly  insoluble  compound,  and  would 
be  of  doubtful  value  from  a  nutrition  standpoint,  amounts  to  about 
20  percent  of  the  total  phosphorus." 


PHOSPHORUS  METABOLISM  99 

PHOSPHORUS  COMPOUNDS  OF  ALFALFA    HAY    AS    AFFECTED    BY 
COMPOSITION  OF  FERTILIZER  (Ames  and  Boltz,  1912) 


Fertilizing     elemen 
per   acre 

ts 

Lime 

Forms    of   Phosphorus 

Total 

Organ- 
ic 

Inor- 
ganic 

Organ- 
ic 
Percenl 
of  total 

Water- 
soluble 
Percent 
of  total 

Phos- 
phorus 

Potas- 
sium 

Nit 

per 
ro-       acre 
n 

in- 
soluble 

First    cutting 
First    cutting 

Lbs.     Lbs.. 

Lbs 

Lbs. 
2,500 

Percent 
.2382 
.2734 

Percent 
.1180 
.1267 

Percent 
.1201 
.1466 

Percent 
49.54 
46.35 

Percent 
80.32 
80.22 

Percent 
.0560 
.0559 

Second    cutting .  . 
Second    cutting.  . 

Unfertilized    .  . 

2,500 
5,000 

.1885 
.2106 

.1190 

.1270 

.0694 
.0836 

63.13 
60.31 

73.24 
65.71 

.0455 
.0519 

First    cutting 
First    cutting 

45 
45 

2,500 
5,000 

.3415 
.3128 

.1494 
.1513 

.1921 
.1615 

43.75 
48.37 

83.50 
86.48 

.0947 
.0656 

Second    cutting.  . 
Second    cutting .  . 

45 
45 

2,500 
5,000 

.2507 
.2447 

.1183 
.1529 

.1324 

.0919 

47.19 
62.49 

79.47 
68.22 

.0550 
.0480 

First    cutting 
First    cutting .... 

45 
45 

25 
25 

2,500 
5,000 

.3116 
.2913 

.1473 
.1295 

.1643 
.1618 

47.28 
44.46 

85.48 
83.92 

.0619 
.0598 

Second    cutting.  . 
Second    cutting.  . 

45 
45 

25 
25 

2,500 
5,000 

.2674 
.2184 

.1402 
.1265 

.1272 
.0919 

52.43 
57.92 

73.80 
77.50 

.0473 
.0529 

First    cutting.  .  .  . 
First    cutting .... 

45 
45 

25 
25 

3 
3 

L2         2,500 
2         5,000 

.2914 
.2388 

.1407 
.1169 

.1507 
.1219 

48.28 
48.96 

78.53 
73.17 

.0640 
.0515 

Second    cutting.  . 
Second    cutting .  . 

45 
45 

25 
25 

] 

2         2,500 
2         5,000 

.2340 
.2316 

.1329 
.1191 

.1011 
.1125 

56.80 
51.43 

70.58 
77.50 

.0445 
.0529 

First    cutting.  .  .  . 
First    cutting .... 

45 
45 

2         2,500 
2         5,000 

.2603 
.2758 

.1158 
.1243 

.1445 
.1515 

44.49 
45.07 

85.87 

.0689 
.0508 

Second    cutting.  . 
Second    cutting.  . 

45 
45 

2         2,500 
2         5,000 

.2423 
.2316 

.1456 
.1344 

.0967 
.0972 _ 

60.10 
58.03 

69.40 
79.46 

.0506 
.0522 

*First    cutting.  .  . 
First    cutting.  .  . 

24 
24 

56 
56 

7 

7 

2         2,500 
2         5,000 

.2472 
.2698 

.1096 
.1248 

.1376 
.1450 

44.34 
46.26 

.0662 
.0522 

*JSecond    cutting. 
Second    cutting. 

24 
24 

56 
56 

7 
7 

2         2,500 
2         5,000 

.2184 
.2232 

.1174 
.144C 

.1010 

.0786 

53.75 
64.79 

::::: 

.0410 
.0543 

*From   16,000   pounds   of  stable   manure. 

Vuaflart  (1911,  1912)  reports  analyses  from  which  it  appears 
that  the  phosphoric  acid  variations  in  flours,  from  year  to  year, 
do  not  correspond  with  those  of  wheats  of  the  same  year.  He  found 
also  that  the  flour  was  not  as  much  enriched  in  phosphoric  acid  by 
increased  superphosphate  treatment  as  was  the  grain. 

PHOSPHORIC  ACID  OF  WHEAT  AND  FLOUR 
Parts  per  100  Parts  of  Nitrogen  (Vuaflart,  1912) 


Wheats 

Flours 

Maximum 

Minimum       Mean 

Maximum 

Minimum 

Mean 

1908 
1909 
1910 
1911 

59.8 
52.3 
56.7 
58.1 

49.8 
42.8 
37.4 
44.2 

54.5 
47.5 
47.4 
49.9 

18.1 

17.9 
16.7 
22.1 

14.1 
12.1 
11.3 
12.8 

15.3 
14.4 
13.7 
17.9 

100 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


Swanson  (1913)  shows  that  the  mineral  constituents  of  flour 
are  much  more  prominently  influenced  by  the  presence  or  absence  of 
bran  than  by  variations  in  the  ash  content  of  the  wheat,  the  phos- 
phorus content  of  the  flour  varying  with  its  grade. 

Forbes,  Beegle  and  Mensching  (1913)  also  studied  the  effects 
of  fertilizers  on  the  phosphorus  of  the  wheat  grain.  The  wheats  rep- 
resented by  the  following  analyses  are  from  the  1911  crop  of  the 
5-year  rotation  series  of  the  Department  of  Soils  of  the  Ohio  Agr. 
Exp.  Station. 

MINERAL  ELEMENTS  OF  WHEATS  VARIOUSLY  FERTILIZED— Parts  per 

100  of  Dry  Substance 


Description 


1  Unfertilized 

2  Phosphorus    

3  Potassium 

4  Unfertilized 

5  Nitrogen     

6  Nitrogen ;    phosphorus 

7  Unfertilized     

8  Phosphorus;   potassium 

9  Nitrogen ;    potassium  .  . 

10  Unfertilized 

11  Nitrogen;    phosphorus; 
potassium     

12  Nitrogen;    phosphorus; 
potassium     

13  Unfertilized     


1 

Inor- 

Ash 

Potas- 

So- 

Cal- 

Magne- 

Sul- 

Chlor- 

Phos- 

ganic 

dium 

cium 

sium 

phur 

ine 

phorus 

phos- 
phorus 

1.87 

.523 

.122 

.051 

.153 

.243 

.089 

.403 

.0229 

1.82 

.561 

.127 

.055 

.149 

.224 

.073 

.406 

.0219 

1.73 

.497 

.135 

.047 

.152 

.237 

.112 

.37 

.0192 

1.71 

.442 

.144 

.049 

.150 

.256 

.100 

.357 

.0181 

1.73 

.467 

.147 

.055 

.154 

.253 

.096 

.349 

.0202 

1.75 

.467 

.139 

.057 

.149 

.243 

.085 

.358 

.0186 

1.80 

.473 

.113 

.056 

.152 

.248 

.102 

.35.6 

.0208 

1.72 

.443 

.157 

.044 

.145 

.213 

.087 

.372 

.0179 

1.61 

.465 

.136 

.043 

.139 

.258 

.100 

.337 

.0185 

1.67 

.459 

.128 

.052 

.144 

.245 

.091 

.340 

.0179 

1.78 

.449 

.154 

.046 

.150 

.228 

.080 

.395 

.0207 

1.77 

.456 

.128 

.048 

.149 

.228 

.079 

.388 

.0176 

1.82 

.451 

.168 

.053 

.149 

.241 

.090 

.359 

.0170 

Organ- 
ic 
phos- 
phorus 


.380 
.384 
.351 
.339 
.329 
.339 
.335 
.354 
.318 
.322 

.374 

.370 
.342 


According  to  Hartwell  (1913)  the  phosphorus  content  of  the 
purple-top,  flat,  or  strap-leaf  turnip  (Brassica  rapa,  L.)  is  most 
markedly  influenced  by  the  amount  of  available  phosphorus  in  the 
soil,  so  much  so  that  he  recommends  its  use  as  a  means  of  determin- 
ing the  relative  amount  of  available  phosphorus  in  different  soils. 
Of  the  range  of  variation  in  the  specific  tests  reported  he  says : — 
"The  percentages  of  phosphorus  in  dry  matter  varied  from  .27  in 
turnip  roots  grown  on  an  extremely  deficient  soil,  to  1.82  in  turnips 
from  a  soil  so  abundantly  supplied  with  phosphorus  that  further 
manuring  even  depressed  the  yield."  The  inorganic  phosphorus 
(Hartwell  and  Hammett,  1911)  was  found  to  be  influenced  much 
more  than  the  total  phosphorus.  Previously  (Hartwell  and  Quantz, 
1910)  it  had  been  found  that  about  70  percent  of  the  phosphorus 
of  this  turnip  is  extractable  with  0.2  percent  HC1,  and  mostly  pre- 
cipitable  from  this  extract  by  molybdate  and  magnesia  mixtures; 
apparently  then,  it  is  present  largely  as  inorganic  phosphorus. 


PHOSPHOKUS  METABOLISM  101 

Summary.  -  All  things  considered,  we  regard  the  observed  ef- 
fects of  soils,  fertilizers,  and  climate  upon  the  phosphorus  content 
of  foods,  in  so  far  as  they  are  derived  from  grains,  to  be  without  im- 
portant practical  significance.  These  variations,  as  a  rule,  are  not 
large ;  the  food  products  of  greatest  value  contain  but  small  propor- 
tions of  the  total  phosphorus ;  and  neither  the  farmer  nor  anyone 
else  is  dependent  for  cereal  foods  upon  the  products  of  any  particu- 
lar farm. 

With  reference  to  roughage,  however,  the  case  is  different; 
roughage  can  not  be  economically  transported ;  it  is  so  bulky.  Prac- 
tically speaking,  the  farmer  is  largely  dependent  on  the  roughage  of 
his  own  farm,  especially  so  for  green  forage.  The  variations  in 
the  phosphorus  content  of  the  roughage  are  large,  and  these  varia- 
tions as  affected  by  soil,  fertilizer  and  climate  constitute  factors  of 
first-class  practical  importance  to  the  breeder  of  live  stock,  espec- 
ially as  affecting  the  growth  of  bone. 

ADDITION  OF  PHOSPHATES  TO  SILAGE 

An  unusual  use  of  a  phosphate  is  reported  by  Sani  (1912).  In 
the  proportion  of  300  gm.  CaH2P04  per  quintal  of  green  clover  this 
salt  preserved  the  foodstuff  in  a  silo  with  much  less  chemical  change 
and  loss  than  occurred  in  untreated  silage.  The  temperature  of 
the  treated  clover  did  not  rise  so  high  as  in  the  untreated  silage, 
and  after  removal  from  the  silo  it  was  found  to  be  immune  to  mold. 
The  flowers  retained  almost  their  natural  color.  In  11  months  the 
treated  fodder  lost  13.7  percent  of  its  weight  and  the  untreated  fod- 
der 18.68  percent. 

Forbes  and  Fritz  (1914)  studied  the  effects  of  the  ensilage 
process  upon  the  solubility  of  rock  phosphate  (floats).  The  phos- 
phate was  added  to  the  green  corn  at  the  time  it  was  put  into  the 
silo.      The  table  on  the  following  page  sets  forth  the  results. 

The  increase  in  total  phosphorus  during  the  ensilage  process 
shows  that  there  was  a  loss  of  8.2  percent  of  dry  substance  from  the 
untreated  corn  and  3.5  percent  from  the  phosphated  corn. 

The  increase  in  water-soluble  phosphorus  in  the  untreated  corn 
was  not  quite  equal  (7.4  percent)  to  the  arithmetical  increase  due  to 
the  loss  of  dry  matter. 

During  the  ensilage  of  this  untreated  corn  there  was  a  loss  of 
citrate-soluble  phosphorus,  in  the  residue  from  water  extraction, 
which  signifies  a  process  of  reversion  to  less  soluble  forms. 

The  one  significant  increase  during  the  ensilage  of  the  untreat- 
ed corn  was  in  the  inorganic  phosphorus  soluble  in  0.2  percent  HC1. 


102 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


This  was  much  more  than  enough  to  account  for  arithmetical  in- 
crease from  loss  of  dry  substance. 

PHOSPHORUS  IN  SILAGE  CORN  WITH  AND  WITHOUT  ADDED  FLOATS, 

AND  IN  SILAGE  MADE  FROM  THE  SAME 

Percent  Water-Free  Basis 


Product 

Total  P 

Water-soluble 
P 

Citrate-soluble 
P     • 

Inorganic     P 
soluble  in 
0.2%    HC1 

Total  water- 
sol.  -+-  citrate- 
sol.  P 

Untreated    green    sil- 

0.200 
0.203 
0.218 
0.207 

0.231 

0.228 
0.214 
0.224 

0.374 
0.371 
0.367 
0.371 

0.405 
0.387 
0.361 
0.384 

0.151 
0.150 
0.147 
0.149 

0.159 
0.161 
0.161 
0.160 

0.135 
0.138 
0.135 
0.136 

0.158 
0.157 
0.155 
0.157 

0.020 
0.021 
0.021 
0.021 

0.009 
0.007 
0.009 
0.008 

0.055 
0.064 
0.065 
0.061 

0.061 
0.061 
0.059 
0.060 

0.086 
0.082 
0.082 
0.083 

0.112 
0.110 
0.112 
0.111 

0.156 
0.181 
0.178 
0.172 

0.234 
0.243 
0.235 
0.237 

Silage    from   untreat- 

o.iess 

G-reen      silage       corn 
plus    floats 
(250:1)     

Silage   from      treated 

0.197 

oiii 

In  the  phosphated  corn  there  was  a  loss  of  water-soluble  phos- 
phorus simply  through  the  addition  of  the  floats,  that  is,  the  water- 
soluble  phosphorus  in  the  fresh  corn  was  0.149  percent  and  in  the 
phosphated  corn  0.136  percent,  which  probably  signifies  a  combina- 
tion of  water-soluble  phosphorus  of  the  corn  with  bases  in  the  floats. 
This  probably  took  place  during  the  partial  drying  at  50°  C,  though 
perhaps  to  some  extent  during  the  subsequent  storage  of  the  sample 
for  nearly  a  year  before  the  analyses  were  made. 

The  water-soluble  phosphorus  in  the  phosphated  silage  was  not 
higher  than  in  the  untreated  silage.  The  excess  of  water-soluble 
phosphorus  in  the  phosphated  silage  over  the  amount  in  the  unen- 
siled,  phosphated  corn  was  more  than  enough  to  account  for  the  loss 
in  dry  matter,  but  was  not  as  great  in  amount  as  in  the  silage  from 
the  untreated  corn,  again  suggesting  reversion. 

The  citrate-soluble  phosphorus  in  the  treated  fodder  and  in  the 
silage  from  the  same  was  naturally  higher  than  in  the  untreated 
corn  and  silage,  since  a  part  of  the  phosphorus  of  the  floats  was 
citrate-soluble.  There  was  no  increase  in  citrate-soluble  phospho- 
rus, however,  in  the  residue  from  the  water  extraction,  during  the 
ensilage  of  the  phosphated  corn. 


PHOSPHORUS  METABOLISM  103 

The  very  considerable  increase  in  inorganic  phosphorus  soluble 
in  0.2  percent  HC1  during  the  ensilage  of  the  phosphated  corn  gives 
us  the  most  significant  figure  of  the  test.  In  the  treated  corn  fod- 
der the  inorganic  phosphorus  soluble  in  0.2  percent  HC1  was  46.4 
percent  of  the  total,  while  in  the  silage  from  the  same  was  61.7  per- 
cent of  the  total. 

It  is  also  of  interest  that  the  phosphated  silage  contained  more 
than  twice  as  much  inorganic  phosphorus  soluble  in  0.2  percent  HC1 
as  the  treated  silage. 

The  total  phosphorus  of  the  floats  was  12.666  percent,  the 
water-soluble  phosphorus  0.0129  percent  and  the  phosphorus  soluble 
in  0.2  percent  HC1  8.721  percent,  all  on  a  water-free  basis. 

Conclusion.  The  ensilage  of  corn  will  render  soluble  in  0.2  per- 
cent HC1  such  an  amount  of  the  phosphorus  of  floats,  added  to  corn, 
as  to  constitute  a  practical  consideration  in  the  feeding  of  livestock. 

REFERENCES  TO  EXTENSIVE  PBESENTATIONS  OF  TOTAL  PHOSPHORUS 
CONTENT  OF  FOODS 

Wolff  (1871,  1880),  Konig  (1903,  1904,  1910),  Albu  and  Neuberg  (1906), 
Schaumann  (1910),  Sherman  (1911). 


104  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


PART  III 

THE  PHOSPHORUS  OF  ANIMAL  BODIES  AND  PRODUCTS 

GENERAL  STUDIES 
THE  PHOSPHORUS  OF  GROWN  MEN  AND  ANIMALS 

Gilbert  and  Posternak  (1905),  in  their  general  discussion  of 
phosphorus  therapy  from  the  standpoint  of  metabolism,  sum  up 
the  total  phosphorus  of  the  human  body  as  being  about  30-40  gm. 
P205  at  birth,  and  1600  gm.  at  middle  life.  This  1600  gm.  is  made  up 
of  about  1400  in  the  skeleton,  130  in  the  muscles,  12  in  the  brain  and 
nerves,  10  in  the  liver,  6  in  the  lungs,  about  4  in  the  blood,  etc. 

Gilbert  and  Posternak  generalize  with  regard  to  the  phosphorus 
distribution  in  meats  in  the  following  way.  Meat  contains  0.467 
percent  P205,  and  of  this  0.274  percent  (or  60  percent  of  the  total) 
is  made  up  of  organic  compounds. 

Lecithin    0.060 

Phosphoearnic  acid 0.039 

Soluble  in  water  but  not  precipitated  by  lime  0.039 

Nuclein     0.008 

Organic  phosphorus  insoluble  in  water  (other 

than   nuclein) 0.128 

The  rest  is  soluble  in  water  and  is  precipitable  by  lime,  and 
hence  is  looked  upon  as  probably  inorganic  phosphate. 

Beaunis  has  compiled  ash  analyses  of  various  animal  tissues 
and  products,  as  in  the  following  table,  from  which  one  may  observe 
the  relative  prominence  of  phosphoric  and  other  oxides  in  these 
parts.  Approximately  half  of  the  ash  of  bone,  muscle,  brain  and 
liver  is  phosphoric  anhydrid.  In  other  parts  the  proportion  of 
phosphorus  is  much  less.  Since  all  of  the  other  elements  reported, 
except  sulphur,  constitute  larger  percentages  of  their  oxides  than 
does  phosphorus  of  its  oxide,  this  method  of  statement  exaggerates 
the  actual  phosphorus  content  of  these  ashes,  as  compared  with  the 
other  elements. 


PHOSPHORUS  METABOLISM 


105 


H 
U 
P 
Q 
O 

►J 

I— I 

*< 
o 

H 

ui 

j  -a 

■«  ° 
a  "B 

O 

o 

I— I- 

H 

«! 

I— I 

o 
(J 

xn 


P 
M 


O  B 

h  a 


•  !~»00**IO 


.COOCOCOCO      .      .1003COCO 


■  r~oo-s*iocM 


•CDTfli— IOCS 


MCOCMC 


COCO  .TTICOt^O 

c— co  •  TtHF-oo*— i 

dto  'Hoood 

i-ICM  CU.-I 


•  oco 
•oicN 


CO     .CO^lO-rH 
CM     'COCOr-ICO 


CD  >-H 


oo    -wmr^coia 
t)!    •dco'ddd 


■  OM 

•odo 


^3 


dJ 


•r1  TO 

OS 


to  r—  10  CO  oo  co  co    .    .-^fcnf-CM 


co  oo  m  co  io  io  ^n 


t^  Oi  CO  CM  CM  O  O 


■OOHTf 
•OOCMO 


oo    .coinoor~co    .    .nooo 


00      •050SCMCMCM 


CM      -CMCMCMOO 


HTflCM 
•i-HCnJth'o 


t-a     .inr— COCMI>- 

OO      «i— (Oit^r- iCO 


00     -TjIr-I^Hr 


•  CMCOt-h 

•Oi-Ir-i 


•  COOOOOiCO-*!      .O-* 

•  COCO-^I-^CMIO      -^HIO 

•ajror-^o'r-^o    •  t-^cM 


■  1OC002  05CM 
•  cdr-Hi— It-HCO 


•  i— IMHO^OO     .00CM 

•  mcMcocMc^m   •  ■— ios 
■^*incdocM04     oo' 


•*     .OiCMCOCO 
t—     ■CO'"*!— CM 


^ 'O^OH 


OS    -locscnio 

»0      'CO-^CTlTti 


•rHCO     -lO 


•a  3 
'E-S 
o  u 

s  a 

.3 


«  °  C  C/3  3  C 


106 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


From  Wolff's  "Farm  Foods,"  Eng.  ed.,  we  quote  the  following 
table,  from  the  work  of  Lawes  and  Gilbert.  The  two  most  im- 
portant factors  determining  the  percentage  of  phosphorus  in 
animals  is  their  relative  development  of  skeleton,  the  tissue  richest 
in  phosphorus,  and  of  fat,  the  tissue  poorest  in  phosphorus.  Of  the 
three  common  meat  animals,  cattle  contain  the  highest  percentage 
of  phosphorus,  and  swine  the  lowest  percentage  of  the  same. 

MINERAL  SUBSTANCES  OF  THE  BODIES  OF  OXEN,  SHEEP,  AND  PIGS 
Lawes  and  Gilbert — Percent,  Live  Weight 


.       Ox 

Fat 
calf 

Sheep 

Pat 
lamb 

Pig 

Half- 
fat 

Pat 

Thin: 

Half- 
fat 

Pat 

Very- 
fat 

Thin 

Fat 

Phosphoric  Acid.. . . 

1.839 
2.111 
0.085 
0.205 
0.146 
0.040 
0.038 
0.087 
0.059 
0.013 

1.551 
1.792 
0.061 
0.176 
0.126 
0.024 
0.033 
0.071 
0.055 
0.006 

1.535 
1.646 
0.079 
0.206 
0.148 
0.021 
0.041 
0.047 
0.063 
0.005 

1.118 
1.321 
0.056 
0.173 
0.120 
0.037 
0.052 
0.037 
0.072 
0.021 

1.199 
1.350 
0.052 
0.168 
0.104 
0.042 
0.035 
0.053 
0.051 
0.020 

1.040 
1.184 
0.048 
0.148 
0.097 
0.034 
0.031 
0.041 
0.044 
0.026 

1.108 
1.240 
0.055 
0.158 
0.129 
0.030 
0.028 
0.049 
0.066 
0.016 

1.126 
1.281 
0.052 
0.166 
0.103 
0.026 
0.039 
0.043 
0.053 
0.012 

1.066 
1.079 
0.053 
0.196 
0.110 
0.022 
0.053 
0.021 
0.056 
0.005 

0.654 
0.636 

0.032 

0.138 

0.073 

0.013 

Sulphuric  Acid.. .. 
Carbonic  Acid.  -  •  • 

0.029 
0.021 
0.043 
0.003 

1 

Francis  and  Trowbridge  (1910)  have  reported  phosphorus  de- 
terminations on  the  different  parts  of  the  carcass  of  eight  beef  ani- 
mals of  different  ages  and  in  various  conditions.  Their  figures  on 
four  animals  representing  extremes  of  age  or  condition  are  quoted 
on  the  following  page.  Moisture,  fat,  ash  and  total  phosphorus  were 
determined.  The  emaciated  and  the  very  fat  steers  contained,  in 
most  of  the  parts,  less  phosphorus  in  the  dry,  fat-free  tissue  than 
did  the  fat  calf  and  the  aged  cow,  the  two  former  perhaps  being  in 
more  abnormal  states  of  nutrition. 

Nerking  (1908b)  has  estimated  lecithin  in  many  animal  tissues. 
For  data  see  next  page.  The  high  content  of  nerve  tissue  in  lecithin 
is  notable,  the  glandular  organs  contain  considerable  amounts,  and 
there  is  evidence  that  under  certain  conditions  the  bone  marrow 
may  serve  as  a  storage  depot  for  this  important  nutrient. 


PHOSPHORUS  METABOLISM 


107 


PHOSPHORUS  IN  THE  DIFFERENT  PARTS  OF  THE  CARCASS  OF  BEEF 
ANIMALS  (Francis  and  Trowbridge,  1910)  Percent 


Steer;  1  yr.  10 

mos.;  emaciated; 

submaintenance 

for  11  mos.1 

Steer;  11  mos.; 
fat 

Steer;  4  yr.  6  mos. ; 
very  fat 

Cow;  7  yr.  6  mos.; 
fat 

Fresh 

Moisture 

and 
fat  free 

Fresh 

Moisture 

and 
fat  free 

Fresh 

Moisture 

and 
fat  free 

Fresh 

Moisture 

and 
fat  free 

0.019 
0.039 
0.137 
0J59 
0.323 

0.183 
0.333 
0.067 
0.109 
0.163 
0.184 
0.173 
0.179 
0.142 
0.168 
0.170 
0.174 

0.116 
0.101 
0.801 
1.037 
2.305 

1.356 
1.299 
0.479 
0.815 
0.762 
0.887 
0.882 
0.844 
0.637 
0.804 
0.824 
0.799 

0.055 
0.072 

lo.'isi 

0.193 
0.347 
0.020 
0.034 
0.164 
0.191 
0.143 
0.158 
0.125 
0.149 
0.163 
0.167 

0.266 
0.225 

i.'isi  -j 

1.204 
1.339 
1.470 
1.072 
0.818 
0.922 
0.968 
0.883 
0.798 
0.821 
0.964 
0.869 

0.021 
0.048 
0.076 
0.117 
0.425 

0.175 
0.307 
0.016 
0.012 
0.142 
0.146 
0.093 
0.098 
0.064 
0.082 
0.123 

0.102 
0.149 
1.756 
0.887 
2.453 

0.953 
1.179 
1.045 
0.710 
0.759 
0.842 
0.892 
0.872 
0.732 
0.804 
0.799 

0.030 
0.072 
0.103 
0.164 
0.354 

0.140 
0.339 
0.016 
0.020 
0.176 
0.182 
0.126 
0.141 
0.125 
0.149 
0.158 
0.153 

0.153 

0.222 

1  772 

1.071 

2.565 

Digestive  and  excretory 

1.115 

1.323 

Offal  fat 

Shin,  shank,  head  and  tail . . 

0.899 
0  983 

1  176 

1.168 

1.018 

Rib 

1.112 

0  923 

Composite  of  leans  and  fats  •  • 

1.085 

(x)  "The  condition  of  the  skeleton  of  this  steer  was  remarkable;  the  marrow  having 
practically  disappeared,  being  replaced  with  a  watery  malodorous  liquid  with  none  of  the 
properties  of  normal  marrow  and  totally  lacking  in  greasy  or  fatty  appearances." 

LECITHIN  DETERMINATIONS   IN   ANIMAL  TISSUES— (Nerking,   1908) 

Percent,  Dry  Basis 


Organs,  etc 


IV 
Rabbit 


V 
Rabbit 


Mean  of 

IV  and  V 

Rabbit 


VI 

Cat 


VII 

Hedgehog 


Lungs 

Heart 

Brain 

Spinal  cord . . . 

Kidneys 

Spleen 

Eyes. 

Stomach 

Liver 

Intestine 

Gall 

Blood.. 

Muscle 

Bones 

Pelt 

Bone  marrow . 

Testes 

Adrenals 

Whole  animal 


5.99 
5.618 
12.23 
33.62 
4.30 
3.27 
3.05 
2.67 
4.22 
0.48 
Trace 
0.914 
2.68 
0.283 
0.568 
4.531 


5.93 
6.108 
12.593 
36.75 
5  73 
5.20 
1.32 
3.94 
3.42 
0.777 

0.'813 

2.49 

0.260 

0.392 

0.89  1 

3.39 

5.54 


5.96 
5.863 
12.41 
35.19 
5.02 
4.24 
2.19 
3.31 
3.82 
0.629 

6!  864 

2.59 

0.272 

0.480 

2.711 


6.10 
4.55 
13.74 
26.20 
6.26 
0.39 


4.99 


5.36 


4.28 
10.49 
22.31 
18.19 
8.55 
6.56 

6\37 

5.23 

1.508 

Trace 

3.71 

0.871 

0.585 

41.7  l 

11.27 

92.00 


0.382  2 


0.4478  2 


0.82142 


Other  rabbits 
1 

n 
m 

1  These  determinations  were  made  on  fresh  substance. 
■  Percent  of  live  weight. 


Lecithin,  percent  of  live  weight 
0.3626 
0.4025 
0.7995 


108  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

THE  PHOSPHORUS  OF  FETUSES  AND  NEW-BORN  YOUNG 

Analyses  of  the  entire  bodies  of  animals  dying  at  birth,  or  very 
soon  after,  have  been  discussed  chiefly  in  connection  with  Bunge's 
suggestion  that  the  composition  of  the  ash  of  new-born  animals  gen- 
erally corresponds  with  that  of  the  ash  of  milk,  perhaps  peculiarly 
with  that  of  the  milk  of  the  same  species  (Bunge,  1874).  Our 
table  shows  the  ash  analyses  of  Bunge  (1874,  1886,  1889)  and  of 
Abderhalden  (1899c)  on  the  young  of  several  animals,  and  those  of 
other  investigators  on  new-born  human  infants.  (Giacosa,  1895; 
deLange,  1897,  1900;  Michel,  1899,  1900;  Hugounenq,  1899b,  1900; 
Soldner,  1902.)  In  this  direction  the  work  of  Soldner,  in  conjunction 
with  W.  Camerer,  Jun.,  is  more  extensive  than  that  of  others,  and 
their  latest  results  expressed  as  total  grams  in  the  body  may  be  of 
interest,  as  given  at  the  foot  of  the  table.  (Our  figures  here  are 
taken  from  Camerer  and  Soldner,  1903.) 


PHOSPHORUS  METABOLISM 


109 


fc 

pel 

O 

CO 

P5 

PS 

£ 

o 
W 

W  H 

SP 

fo 

-sd 

O  a 

nrt 

P 

P 

o 

o 

PS 

PQ 

> 

W 

JH 

PS 

PU 

h- 1 

H 

Q 

fc 

W 

W 

H 

PS 

W 

o 

w 

Pu 

H  W 

fe 

PS 

O  GQ 

co 

<1 

fc 

CO 

O   M 

1— 1 

H 

^ 

S 

k; 

^ 

S 

<j 

PS  Q 

w 

fc 

H 
H 

<s 

fi 

K-l 

^ 

<J 

<1 

PS 

fe 

1— 1 

0000O3lC31>-COOiCO00CO 
i-Ht— IrHiHi— irH»-4e4tHCO 

.      .     .K3MH 

•    •    -mm   ..... 
■    '    -o'o   •  •    • 


iococnj^^hooouocmco 

Ni-HQOlOOTflr-HOOOt- 
CN1  >-i  CM  CM 0  i-I  H  cm  00  ■* 


COt^t--0-# 

HO     .IOCOCOCOOCOOO 

com   'Hnoohho 
o'o   •oo'ooooo 


COt-HOIOOOOCM'— <-HCM 
COTf*TJ1tncOCOTJ<lOTj<CM 

OOOOOOOOOrH 


in-HmoKMOiwooH 

C~U30iC0rHlOT*C0COTtH 
CO  i-4  CO  Tli  O  OS  OS  i-H  CD  1-5 


ISO     .O-*C003CD00O 
COCO     -OOCOCMIOCO-* 


cocm    -cocMr-icMcMiMCM 


001(      .OJt—  1^-0500  CO  CO 
0OO5     -CM00O5r~COIO00 

o>-I    'oir-IcNCNiojoioi 


IOI^CTiC5CMTt(^*^<0303 
01C»HOONf-t^»01 
00  00     „CJS  OS  OO  00  00  00  00 


l>.COCMCOi— f-^HCMT^lOCO 

r^coi^-cMcoosi— icoco-^i 
»o  CO  id  "*  cd~*  t^  t^  00  OS 


IO»— <~H00CO^*COT*ICMOi 
COCOOCMCOOiCMOOTjft^. 

t^t^00*U0I>^T-HO03C3i— H 

COCOCOCOCO-'SI'^COCO^ 


OiOi        05COCO-#-*CM-* 
COCO       CO  00  CM  CM  CO  F- CM 


ooooooo 


Ot^Oi-ICOOiCMr-IO»Tt( 
THCOCMmTfl^-llOCOOO-Vl 


T-iOOll 


CMQ3O0C00CM 
OiOOCO^CNOO 

r-J  00  i-i  ©  00  HO  TK  1T5  CT>  CM 

TflCOTflTKCOCOCOCOCMCO 


COO      .<Mr~CO00rHT*O5 
CMOO      •!— I  COOS  CM  CM  CO  t>- 

ood    ■oo'r-irsododocd 


Or«     .OCOtSf. 105CM05 


CM  CO 


CMOOOf— 1TJHTJ1© 

co'  C-i  o'  ©  00  i-i  oo 


W03020CM*J<Tj<'^a2CD 
fflHnoOt;r-NC0  3) 
00      .     „03  03  CO  00  00  OO  00 


rtd— *o)Oe0coa;cj'.H 
g^Sdjibobohoejirt 

o   Kg  5 


d  d  d  d  d  +j 

td  rt  rt  nj  ctj  *jj    . 

333a3drtoo3 

WWWWMKOOQO 


-.  t»     d  rt  •§ 

g  rt  «  d_.  bo  bo  ho  ho  rt 

Sj-g  g-g  d  d  d  dfl 

5«S^WWMn's 

!o 

CO 


5* 

be 


o 
PQ 


bo 

u 

> 


d  d  d  d  d 

rt  ri  ri  7i  rt 

aaaaag 

3  3  3  3  3rt"5  g'o'a 

WWWMM«Ofiqci 


ho  ho-9 


a  *  * 

O     fr)    «H 

5    ft  2 


rtf     TH 


I  8  .a 


rt         -^  w 


110 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


Michel  (1899,  1900)  gives  two  reports  of  the  same  analyses  of 
human  embryos  and  fetuses  of  different  stages,  and  of  a  new-born 
infant.  We  quote  the  table  which  gives  these  analyses  on  the  basis 
of  100  gm.  of  fresh  substance.  These  data  reflect  the  rapid  deposit 
of  phosphorus  which  occurs  late  in  fetal  life. 

COMPOSITION    OF    HUMAN    FETUSES    AND    A    NEW-BORN    INFANT 

Michel  (1899) 


Body 
weight 

Grams 

Percents  of  fresh  substance 

fetus 
Months 

Water 

Nitrogen 

Protein 
(calcu- 
lated) 

Ash 

CaO 

MgO 

P2O5 

CI 

Fat 

2.5 

3.4 

5 

5 

6 

7 

Pull  term 

17.80 

125.80 

445 

448 

672 
1024 
3335 

93.82 
89.95 
87.80 
86.95 
85.02 
84.73 
69.16 

0.685 
1.100 
1.322 
1.390 
1.644 
1.563 
2.179 

4.39 
7.05 
8.46 
8.90 
10.50 
10.04 
13.96 

L729 
1.948 
2.485 
2.512 
2.487 
3.37.3 

6!465 
0.597 
0.790 
0.850 
0.804 
1.393 

0.027 

0.0258 

0.0314 

0.0328 

0.0307 

0.0405 

0.489 
0.643 
0.842 
0.833 

0.788 
1.282 

6.'240 

0.289 
0.193 

6 '.397 
0.888 
0.798 
1.210 
1.823 
11.750 

Brubacher  (1890),  in  his  study  of  rachitis,  gives  the  following 
mineral  determinations  on  the  entire  bodies  of  two  undernourished 
human  fetuses. 

BRUBACHER'S  ANALYSES  OF  HUMAN  FETUSES 
Percents  of  Fresh  Substance 


Age   of   fetus 
Weeks 

Water 

Fat 

Ash 

CaO 

MgO 

P2O5 

Si02 

Fe203 

28 
36 

80.75 
75.28 

3.95 

8.42 

3.00 
3.12 

1.04 
1.13 

0.04 
0.04 

1.09 
1.15 

0.01 
0.01 

0.01 
0.01 

Siwertzow  (1909)  presents  the  following  data  on  the  lecithin 
content  of  the  organs  of  human  fetuses  and  children.  He  concludes 
that  of  the  brain,  liver,  heart  and  muscles,  the  brain  contains  more 
lecithin  than  all  these  other  organs  put  together,  the  liver  coming 
next,  and  then  the  heart,  the  muscles  having  least  of  all.  The  lec- 
ithin content  of  each  of  the  organs  gradually  increases  in  the  fetus 
to  a  maximum  at  the  time  of  birth.  During  the  first  months  of 
extrauterine  life  the  lecithin  remains  constant  or  even  decreases, 
and  then  rises  again,  reaching  a  new,  permanent  maximum  at  the 
second  year.  The  facts  established  seem  to  the  author  to  show 
that  in  the  new-born  child  there  is  a  store  of  lecithin,  as  has  been 
shown  for  iron,  and  that  this  is  gradually  used  up  during  the  first 
four  months. 


PHOSPHORUS  METABOLISM 


111 


LECITHIN  CONTENT  OF  ORGANS  OF  THE  HUMAN  FETUS  AND  OF 
YOUNG  CHILDREN  (Siwertzow,  1903)  Percent,  Dry  Substance 


Age 

Number     ex- 
amined 

Brain 

Liver 

Heart 

Muscle 

6    months 

2 

9.39 

0.25 

0.00 

1.30 

8 

1 

11.95 

2.82 

2.22 

1.32 

9 

2 

12.86 

3.80 

3.55 

1.38 

10          " 

4 

16.21 

4.90 

4.51 

2.18 

Child   . 

1      month 

4 

15.66 

3.05 

2.40 

1.54 

2    months 

2 

15.35 

3.20 

2.39 

1.44 

3 

2 

14.85 

2.74 

2.44 

1.65 

4 

2 

16.40 

3.37 

2.13 

1.58 

6           " 

1 

17.67 

4.83 

2.59 

1.83 

10 

1 

21.59 

5.77 

3.93 

2.40 

2       years 

1 

22.78 

8.15 

7.52 

3.86 

3   yrs.    5   mos. 

1 

23.15 

7.58 

7.23 

4.34 

THE  PHOSPHORUS  OF  INFANTS'  BODIES  AFTER  DISEASE 
Steinitz  (1904)  made  analyses  of  the  bodies  of  infants  that  had 
suffered  serious  nutritional  disturbances  and  found  them  not  to  dif- 
fer much,  save  in  fat,  from  the  new-born  infants  reported  by  Cam- 
erer  and  Soldner.  We  give  the  ash  analyses,  together  with  those 
of  Steinitz  and  Weigert  (1904,  1905)  on  a  one-year-old  child  that 
had  died  of  tuberculosis  with  rickets. 

ASH  ANALYSIS  OF  BODIES  OF  INFANTS— Percent  of  Ash 


Case 


I 

II 

III 

IV 


K20 

Na20 

CaO 

MgO 

Fe203 

P2O5 

CI 

8.3 

8.7 

37.8 

1.1 

1.0 

39.8 

6.5 

7.2 

7.7 

38.1 

1.2 

1.2 

38.7 

6.1 

6.85 

7.8 

37.8 

1.32 

0.94 

36.3 

5.7 

6.88 

9.03 

35.85 

0.99 

0.425 

34.7 

7.38 

Cause    of   death 


Nutritional    disturbance 


Tuberculosis 


Cases  I,  II  and  III,  Steinitz   (1904) 
Case  IV,  Steinitz  and  Weigert   (1904) 

THE  PHOSPHORUS  OF  BONES,  TEETH,  MARROW  AND  CARTILAGE 

EEFEEENCES  TO  OLD  WOEK 

Probably  the  most  extensive  of  the  early  work  on  this  subject 
was  that  of  von  Bibra  (1844)  who  published  a  book  of  430  pages  on 
chemical  studies  made  by  himself  and  several  others  on  the  bones 
and  teeth  of  men  and  other  vertebrates.  Other  more  or  less  com- 
plete analyses  are  those  of  Frerichs  (1842),  Nasse  (1842),  Stark 
(1845),  Fremy  (1855),  von  Recklinghausen  (1858),  Edwards 
(1860),  Zalesky  (1866),  Papillon  (1870,  1870-71,  1873),  Volkmann 
(1873),  Mallet  (1874),  Hofmeister  (1873),  and  Siedamgrotsky  and 
Hofmeister  (1879).  From  Fremy's  work  we  quote  the  table  on  the 
following  page. 

Schrodt  (1876)  presents  analyses  of  the  individual  bones  of  a 
normal,  healthy  dog.  The  inorganic  matter  of  the  water-free  bones 
varied  mostly  between  60  and  70  percent,  but  this  figure  for  the 
sternum  was  as  low  as  49.57  percent,  and  in  the  third  neck  vertebra 
as  high  as  72.98  percent.       In  the  ash  the  C02  varied  between 


112 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


4.61  and  9.06  percent,  the  CaO  between  50.40  and  53.28  percent,  the 
MgO  between  0.78  and  1.08  percent  and  the  P205  between  39.08  and 
40.47  percent.  The  ratio  of  calcium  carbonate  to  calcium  phos- 
phate is  said  to  be,  in  compact  bones,  1:6.4,  and  in  spongy  bones 
1:5.7.  It  is  noted  that  the  ash  constituents,  with  the  exception  of 
carbonic  acid,  occur  in  almost  the  same  quantities  in  relation  to  one 
another  in  all  the  bones. 
ANALYSES  OF  BONE  OF  VARIOUS  ANIMALS   (Fremy,  1855)— Percent 


Animal 


Bone 


Woman   

Man     

Egyptian  iaummi 

Dog 

Lion,  young   .  .  . 

Sea-lion     

Rabbit     

Elephant,  India 
Rhinoceros,  Java 
Calf,    5   mos.    .  . 

Cow,  old 

Ox 

Bull    

Lamb     

Sheep    

Goat    

Sperm  whale    .  . 

Eagle     

Owl      . 

Fowl    

Turkey    

Heron    

Teal     

Sea  turtle 

Crocodile    

Carp    

Eel     

Ray 

Cod    .'.[\\\'.'.\'. 


Skull 
Femur 


Femur 


Humerus 
Femur 


Ash 


Carapace 
Bony  skin 


Cartilage 
Spines 


64.1 
64.2 
65.0 
62.1 
64.7 
63.1 
66.3 
66.8 
65.3 
69.1 
71.3 
70.4 
69.3 
67.7 
70.0 
68.0 
62.9 
70.5 
71.3 
68.2 
67.7 
70.6 
73.5 
64.0 
64.6 
61.4 
57.0 
30.0 
65.3 
61.3 


Calcium    phos- 
phate 


57.8 
56.9 
58.7 
59.0 
60.0 
53.9 
58.7 
62.2 
60.0 
61.2 
62.5 
61.4 
59.8 
60.7 
62.9 
58.3 
51.9 
60.6 
61.6 
64.4 
63.8 
62.5 
68.4 
56.0 
58.3 
58.1 
56.1 
27.7 
64.4 
55.1 


Magnesium 
phosphate 


1.7 
1.3 
1.7 
1.2 
1.5 
1.5 
1.1 
1.2 
2.3 
1.2 
2.7 
1.7 
1.5 
1.5 
1.3 
1.2 
0.5 
1.7 
1.5 
1.1 
1.2 
1.5 
1.3 
1.2 

trace 
1.1 

trace 


1.3 


Calcium      car- 
bonate 


10.9 

10.2 
5.9 
6.1 
6.3 
9.3 
6.3 
5.6 
5.2 
8.4 
7.9 
8.6 
8.4 
8.1 
7.7 
8.4 

10.6 
8.4 
8.8 
5.6 
5.6 

10.2 
5.6 

10.7 
9.7 
4.7 
2.2 
4.3 
1.3 
•    7.0 


COMPOSITION  OF  THE  BONES  OF  A  GANDER  (Hiller,  1885) 

Bone 

Inorganic 

matter   of 

fat-  and 

water-free 

bones 

Percent 

Percent    of    ash 

Carbonic 
acid 

Lime 

Magnesia 

Phos- 
phoric 
acid 

58.48 
56.64 
54.99 
47.94 
62.67 
60.61 
56.72 
57.11 
59.45 
58.91 
67.16 
65.91 
61.75 
61.13 
62.50 
61.54 
55.71 

4.67 
5.30 
4.31 
4.03 
5.20 
5.36 
4.00 
5.67 
4.34 
4.48 
4.93 
5.49 
4.48 
4.92 
5.47 
5.25 
5.35 

51.83 
51.61 
52.16 
51.52 
51.57 
52.04 
52.42 
51.86 
52.88 
52.18 
52.50 
52.11 
52.39 
51.30 
51.79 
51.66 
51.34 

0.68 
0.77 
0.83 
0.92 
0.85 
0.67 
0.75 
0.80 
0.67 
0.81 
0.81 
0.84 
0.84 
0.76 
0.86 
0.85 
0.75 

38.90 

2 

38.83 

3 

39.08 

4 

38.86 

5 

39.54 

g 

38.63 

7 

38.11 

8 

Ribs    

38.19 

9 

39  45 

10 

38.48 

11 

38.75 

12 

13 

Right  and  left  radius  and  ulna 

38.57 
39.28 

14 

38.97 

15 
16 

38.50 
38.99 

17 

39.53 

Hiller  (1885)  made  a  similar  study  (though  with  less  separa- 
tion of  the  individual  bones)  of  the  bones  of  a  two-and-a-half -year- 
old  gander  in  normal,  average  condition.      The  conclusions  were  the 


PHOSPHORUS  METABOLISM 


113 


same  as  those  of  Schrodt,  except  that  the  bone  showing  the  highest 
content  of  inorganic  matter  was  the  humerus,  and  the  ratio  of  cal- 
cium carbonate  to  calcium  phosphate  varied,  independently  of  the 
compactness  or  sponginess  of  the  substance,  between  1 :6  and  1 :8. 

ANALYSIS  OF  MINERAL  MATTER  AS  OBTAINED  BY  REMOVAL  OF  ORGANIC  MATTER 
"  WITH  GLYCERIN  AND  POTASSIUM  HYDRATE 

Gabriel  (1894)  preferred  obtaining  the  mineral  matter  of  bones 
and  teeth  by  removing  the  organic  matter  by  heating  the  powdered 
bone  with  alkaline  glycerin  at  200°C.  The  substance  obtained  in 
this  way  retains  water  and  carbonic  acid  which  are  driven  off  at  the 
high  temperature  of  ashing.  Gabriel  says  that  the  mineral  matter 
of  bones  contains  water  in  two  forms,  water  of  crystallization,  and 
water  of  constitution,  the  latter  not  being  driven  off  by  heat  alone 
but  only  by  heating  with  silicic  acid. 

COMPARISON  OF  THE  MINERAL  MATTER  OF  THE  TEETH  OF  CATTLE 

AS  PREPARED  BY  ASHING  AND  BY  THE  GLYCERIN  METHOD 

Gabriel  (1894)— Percent  of  Ash 


Treatment 

CaO 

Mg-O 

K2O 

Na20 

H20of 
crystal- 
ization 

P2O5 

CO2 

CI 

1 

Organic   matter   removed  by 

50.68 

50.76 
53.67 

50.59 

1.52 

1.52 
1.56 

1.47 

0.23 

0.20 
0.25 

0.24 

0.97 

1.16 
1.13 

1.07 

2.27 
2.21 

2.21 

38.78 

38.88 
41.55 

39.13 

4.16 

4.09 
0.59 

4.09 

0.05 

2 

Organic   matter   removed  by 

fr.05 

3 

4 

Ashed  in  the  usual  way 

(3)  computed  to  H2O  and  CO2 

0.10 
0.09 

ANALYSES     OF     "GLYCERIN-ASHES"     OF     BONES     OF     DIFFERENT 
SPECIES    (Gabriel,  1894)— Percent  of  Ash 


H20    of 

Animal 

Bone 

CaO 

MgO 

K20 

Na20 

crystal- 
lisation 

Pa06 

co2 

CI 

Teeth 

50.76 

1.52 

0.20 

1.16 

2.21 

38.88 

4.09 

0.05 

Cattle    

Tooth,    enamel 

51.98 

0.53 

0.20 

1.10 

1.80 

39.70 

3.23 

0.21 

Cattle    

Tooth,  dentine 

50.36 

1.83 

0.14 

0.80 

2.90 

38.60 

3.97 

0.03 

Humerus 

51.31 

0.77 

0.32 

1.04 

2.46 

36.65 

5.86 

0.01 

Cattle    

Femur 

51.28 

1.05 

0.18 

1.09 

2.33 

37.46 

5.06 

0.04 

All    bones 

51.01 

1.27 

0.19 

1.11 

3.05 

38.19 

4.11 

0.06 

INFLUENCE  OF  AGE  ON  THE  COMPOSITION  OF  BONES 

The  changes  in  the  appearance  of  bones  as  an  effect  of  age  are 
so  characteristic  that  anyone  who  is  acquainted  with  beef  recognizes 
at  once  the  soft  vascular  bones  of  young  cattle  and  the  white,  flinty 
bones  of  old  animals.  Cattle  from  regions  where  they  are  not  mar- 
keted at  an  early  age  are  sometimes  distinguished  from  cattle  raised 
in  more  intensive  farming  regions  by  referring  to  the  former  as 
"hard-boned"  cattle. 


114 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


Such  data  as  we  have  on  the  changes  in  composition  are  all  old, 
but  probably  they  give  a  fair  interpretation  of  the  facts.  They 
are  from  Weiske  and  from  Wildt  (Weiske,  1872a,  1889;  Wildt, 
1872).  With  both  the  birds  and  the  rabbits  which  Weiske  exam- 
ined the  water  content  of  the  bones  decreased  as  the  animals  in- 
creased in  age,  and  in  the  dry  bones  the  mineral  content  increased 
and  the  organic  content  decreased.  The  fat  content  of  the  water- 
free  bones  increased  in  the  rabbit,  while  in  the  birds  it  is  said  to  be 
larger  in  the  young  than  in  the  old  fowl.  With  regard  to  the  com- 
position of  the  ash,  we  have  computed  averages  from  Weiske's  rab- 
bit study. 

LIME,  MAGNESIA,  AND  PHOSPHORIC  ACID  CONTENT  OF  THE  ASH  OF 
RABBIT  BONES  AT  DIFFERENT  AGES  (Weiske,  1872)  Percent 


Age 

CaO 

MgO 

P206 

51.24 
53.45 
53.68 
54.06 

1.60 
1.22 
1.23 
1.07 

42.39 

42.47 

41.53 

Calcium  increased,  magnesium  decreased  and  phosphorus  re- 
mained nearly  constant  in  the  ash.  The  same  relations  held  in 
Weiske's  data  on  bird  bones. 

Weiske  (1889)  analyzed  the  bones  of  fowls  of  various  ages,  all 
receiving  grain  food.  These  results  show  with  increasing  age  an 
increase  of  all  the  mineral  constituents  in  the  dry,  fat-free  bone. 
In  the  ash,  there  were  increased  percentages  of  CaO  and  C02  and  de- 
crease in  MgO. 

COMPOSITION  OF  THE   BONES   OF   FOWLS   AS   AFFECTED   BY  AGE 
Weiske  (1889)  Percent,  Dry,  Fat-free  Bone 


Age 

Mineral 
matter 

P2O5 

CaO 

MgO 

Just  hatched    

25.29 

32.77 

37.40 

42.59 

44.92 

42.27 

46.57    ' 

40.12 

48.71 

57.14 

54.20 

57.05 

9.31 

13.45 
14.84 
17.19 
17.79 
16.43 
17.48 
14.70 
18.25 
22.25 
19.98 
21.73 

10.57 
15.35 
17.68 
20.42 
21.97 
20.68 
23.32 
19.99 
24.47 
28.98 
27.86 
28.99 

1   week     

0.36 

2  weeks    

0.47 

4  weeks    

0.54 

0.63 

0.62 

0.49 

24  weeks    

0.49 

0.33 

0.40 

0.61 

0.43 

0.58 

■ MM 

Wildt  (1872)  reported  analyses  of  the  ash  of  the  bones  of  rab- 
bits of  various  ages.  The  following  table  is  from  this  work.  The 
inorganic  substance  increased  with  age  as  also  did  the  calcium  and 
the  carbonic  acid,  while  magnesium  and  phosphorus  diminished. 


PHOSPHORUS  METABOLISM 


115 


ANALYSES  OF  THE  ASH  OF  THE  BONES  OF  RABBITS  OF  DIFFERENT 

AGES  (Wildt,  1872) 


Inorganic  sub- 
stance   x 

Percent 

Percent  of  ash 

Inorganic      sub- 

Age    of   rabbits 

Carbonic 
acid 

CaO 

MgO 

Phosphor- 
ic   acid 

bone 
Percent 

8   months    

53.39 
50.82 
55.18 
58.94 
65.63 
67.68 
68.72 
70.26 
71.77 
74.24 
72.90 
73.65 

3.65 

3.84 
3.99 
4.00 
4.52 
4.69 
4.92 
4.94 
5.54 
5.71 
5.81 
5.66 

52.17 
52.16 
52.10 
51.91 
52.10 
52.49 
52.60 
52.64 
52.78 
52.61 
52.76 
52.84 

1.38 
1.36 
1.26 
1.22 
1.09 
1.01 
1.02 
1.05 
0.93 
0.91 
0.93 
0.83 

42.05 
42.13 
42.19 
42.20 
41.64 
41.03 
40.80 
40.80 
40.05 
40.04 
39.78 
39.80 

15.56 
17.23 
18.62 
23.39 
30.13 
30.90 
37.17 
41.80 
39.22 
44.39 
41.68 
45.00 

(*)      Apparently  reckoned  on  residue  from  water-extraction  of  fat-free  bone. 

See  also  Graff  enberger  (1891). 


COMPOSITION  OF  THE  TEETH 

Hoppe-Seyler  (1862)  submits  a  table  of  analyses  (p.  117)  of 
the  enamel  of  the  teeth  of  several  species  of  animals.  General  sim- 
ilarity is  evident. 

Kiihns  (1895)  compares  human  teeth  at  different  ages.  The 
more  notable  differences  are  in  the  calcium  and  magnesium  of  the 
enamel,  the  former  increasing  and  the  latter  decreasing  with  age, 
and  the  increase  with  age  of  the  phosphorus  of  the  dentine.  (P.  116.) 

Bertz  (1899)  says  that  in  teeth  the  bases  were  found  to  exceed 
the  acids ;  that,  contrary  to  the  findings  of  others,  no  alkalis  or  car- 
bonates were  found ;  that  there  is  no  chlorine  in  enamel ;  that  it  is 
doubtful  if  there  is  a  trace  of  chlorine  in  true  dentine;  and  that 
dentine  is  found  to  contain  twice  as  much  magnesia  and  half  as 
much  fluorine  as  enamel.      (See  table  on  next  page.) 

St.  Bondzynski  and  Gonka  (1907)  found  that  the  organic  sub- 
stance of  calf  teeth,  entirely  freed  from  mineral  matter,  contained 
0.37  percent  phosphorus.  The  ground-substance  of  bone  (femur 
of  horse)  similarly  treated  showed  no  organically  combined  phos- 
phorus. 

For  other  analyses  of  teeth  see  Gabriel,  p.  113,  Gassmann 
(1908)  and  C.  Cohn  (1889),  under  Caries,  and  also  Weiske  (1891b, 
1892, 1894, 1895c,  1896, 1897),  Graffenberger  (1891),  Carnot  (1892) 
and  Aeby  (1873c). 


116 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


o 

CO 

Q 

O 

(— i 

tt 
q  in 

rH   °° 

«s|  w 

i_i     OB 

fifl 

<! 
& 

w 

o 
o 

I-H 

o 
u 


gflS 


r3  rt*» 


WlfllO 

odd 


O)  CO  r-l 
OOOJ 

IMlMlM 


«o$ZiM 


w 

H 
W 

w 

H 

u 

Q 
55 

P 

a 
o 

DO 
H 


0) 

o 

a 

03 

.3 
at 

>> 

«H 

o 

"3 

o 
o 
r-i 
<& 
Ph 

s 

H 
"*.    '. 

d  ; 

o 

0Q 

00 

t-   • 

CO     • 

d   ] 

o 
P4 

rjl 

m   . 

N     • 

d   * 

CO 

o 

oil 

oo 
o    • 

ri   '. 

o 

C3 

o 

© 

CO    • 
r-l     . 

oo'    • 

CO     * 

'3 

03 
U) 
U 

o 

lO     • 

r-l     • 

0>     '. 

Total    dry 
substance 

Percent 

r-l     • 
r-l     • 
00     . 

d   * 
o 

•a  i 


"o  P* 


Ph 


OIH 
CO  t- 

ot- 

rid 


OS  r-l 
CJCO 

do 


cam 

coio 

t-Ol 

do 


(MtJI 
lOTtl 


C0  5O 
Olt- 

rtoi 

eo'cd 


Wd 


PHOSPHORUS  METABOLISM 


117 


COMPOSITION  OF  THE  ENAMEL  OF  TEETH 
Hoppe-Seyler  (1862)  Percent  of  Inorganic  Matter 


Animal 


Man    

Man 

Man    

Hog    

Hog    

Rhinoceros  . 
Elephant  . . , 
Mastodon  .  .  . 
Paleotherium 

Horse 

Dog    


Age 


New-born 
New-born 
New-born 
Young 
Adult 
Fossil 
Fossil 
Fossil 
Fossil 


P205 

CaO 

MgO 

CI 

30.53 

41.42 

0.72 

Trace 

35.69 

44.91 

0.79 

0.15 

36.61 

45.03 

0.50 

t 

39.06 

48.67 

0.74 

0.30 

40.59 

51.57 

0.91 

0.40 

40.57 

51.23 

0.75 

0.42 

38.85 

49.71 

0.92 

0.28 

39.62 

52.82 

0.30 

0.38 

40.20 

52.39 

0.59 

0.37 

40.22 

51.10 

0.56 

0.43 

43.63 

51.46 

2.27 

0.51 

Fe203 


0.34 

Trace 

0.48 

0.47 

1.30 

0.29 

0.17 

0.40 

Trace 


PHOSPHORUS   COMPOUNDS   OF   BONE 

The  Inorganic  Phosphate.  From  all  of  these  analyses  it  is  quite 
evident  that  lime  and  phosphoric  acid  are  the  principal  constituents 
of  the  mineral  part  of  bones,  the  two  together  forming  fully  90 
percent  of  the  ash.  Carbonic  acid  and  magnesia  are  also  certainly 
present,  and  there  is  no  doubt  that  these  are  united  as  some  form  of 
calcium  phosphate,  magnesium  phosphate  and  calcium  carbonate. 
There  are  also  small  quantities  of  sodium,  potassium,  chlorine  and 
fluorine. 

Papillon  (1870,  1870-71,  1873)  claims  to  have  introduced 
strontium,  magnesium  and  aluminum,  and  H.  Stoelzner  (1908) 
strontium,  into  bones  in  place  of  a  small  part  of  the  calcium. 

Attempts  have  been  made  to  assign  definite  formulae  to  the 
calcium  phosphate  and  to  a  combination  of  phosphate  and  carbonate 
of  calcium ;  but,  so  far  as  we  know,  none  of  these  have  been  on  any 
other  basis  than  the  percentage  composition.  Other  chemical  and 
physico-chemical  evidence  .should  be  brought  out  before  confidence 
is  placed  in  any  formula  claiming  to  represent  chemical  union  of  the 
elements  in  the  usual  sense. 

The  articles  in  which  we  have  noted  that  attention  was  given  to 
this  subject  are:  Berzelius,  1816;  von  Gohren,  1865;  Aeby,  1872a, 
1872b,  1873a,  1874a,  1874b;  Wibel,  1874;  Schrodt,  1876;  Levy, 
Moritz,  1894;  Gabriel,  1894 ;  Gassmann,  1910, 1913. 

Gabriel's  conclusion,  after  his  studies  on  the  mineral  constitu- 
ents of  bones  and  teeth,  the  organic  matter  being  removed  by  the 
glycerin  method,  which  avoids  high  temperatures,  he  expresses  in 
this  way:   "The  composition  and  properties  of  bone  and  tooth-ash 


118  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

have  their  simplest  expression  in  the  formula  (Ca3(P04)2-(-Ca5HP3 
013+Aq),  in  which  2 — 3  percent  of  the  lime  is  replaced  by  magnesia, 
potash  and  soda,  and  4 — 6  percent  of  the  phosphoric  acid  by  car- 
bonic acid,  chlorine  and  fluorine." 


Gassmann  favors  the  formula 


OP03Ca 


)Ca 


C03, 


\  OPOaCa 

(which  he  attributes  to  A.  Werner)  on  account  of  having-  found  the 
ratio  of  the  three  constituents  constant  in  this  proportion  in  normal 
and  in  rachitic  bones,  and  from  having  isolated  from  an  ignited  mix- 
ture of  ground  teeth  and  calcium  chloride  a  compound  corresponding 
in  its  chemical  analysis  to  the  analogous  chloride.  To  account  for 
the  increased  proportion  of  Mg  in  rachitic  bones  Gassmann  suggests 
that  this  does  not  belong  to  a  salt-forming  body. 

Lecithin  and  Phosphorus  Determinations  on  Bone  Marrow.  A 
part  of  the  fat  of  bone  marrow  is  phosphorized,  and  on  the  older  un- 
derstanding, which  called  all  the  phosphatids  lecithin,  and  computed 
the  amount  of  lecithin  from  the  percent  of  phosphorus  found  in  the 
alcohol-ether  extract,  lecithin  determinations  on  bone  marrow  are 
reported.  Otolsky  (1906)  and  Bernazky  (1908)  have  studied  in  par- 
ticular the  composition  of  the  lecithin  of  bone  marrow.  From  these 
articles  we  have  seen  only  abstracts.  One  kilogram  of  bone  marrow 
yielded  by  Otolsky's  process  1.3-1.5  gm.  lecithin,  containing  3.25  per- 
cent phosphorus,  and  yielding  choline,  glycerophosphoric  acid  and 
fatty  acids  on  cleavage. 

Bernazky  considered  the  distribution  of  phosphorus  in  the 
organs  of  the  horse,  and  made  a  special  study  of  the  lecithin  of  bone 
marrow.  He  says  that  the  red  bone  marrow  of  the  horse  contains 
twice  as  much  lecithin  as  the  white  bone  marrow,  and  that  no  dif- 
ferences were  observed  in  either  the  amount  or  the  composition  of 
lecithin  in  the  bone  marrow  of  horses  immunized  against  diphtheria, 
scarlet  fever  and  the  yellow  staphylococcus. 

Hutchison  and  MacLeod  (1901-2)  studied  the  red  marrow  from 
the  ribs  of  the  horse.  They  report  that  the  fat,  lecithin  and  choles- 
terin  together  constituted  17.9  percent  of  the  marrow,  the  soluble 
salts  2.34  percent  and  the  insoluble  salts  0.66  percent.  Among  the 
soluble  salts  was  included  0.48  percent  P,05.  A  nucleoproteid  was 
found. 

Nerking  (1908a)  found,  of  lecithin,  in  red  bone  marrow,  0.1576, 
0.1469,  and  0.3006  percent,  and  in  yellow  bone  marrow,  0.1636  and 
0.2046  percent. 


PHOSPHORUS  METABOLISM 


119 


Glikin  (1907,  1908a,  1908b,  1909a)  made  a  number  of  Fe203» 
P206  and  lecithin  determinations  on  the  fat  of  the  bone  marrow  of 
different  kinds  of  animals  at  different  ages  which  seem  to  support 
the  view  that  the  marrow  of  very  young  animals  is  richer  in  lecithin 
than  that  of  older  animals,  and  that  it  is  highest  in  the  young  of 
those  species  which  are  most  helpless  at  birth. 

PHOSPHORUS,  IRON  AND  LECITHIN  DETERMINATIONS  ON  THE  FAT 
OF  BONE  MARROW  OF  SEVERAL  SPECIES  OF  ANIMALS  AT 
DIFFERENT  AGES  (Glikin,  1907)  Percent 


Animal 


Cattle. 
Cattle. 
Cattle. 
Cattle., 
Calf.... 
Calf.... 
Calf.... 
Calf.... 
Horse. . 
Horse  •  • 
Horse . . 
Horse.. 
Horse. . 
Horse.. 


Swine. 
Swine . 
Swine. 
Swine . 
Pigs.. 
Pigs . . 
Pigs1.. 
Pigs2.. 
Pigs. . 
Sheep. 
Sheep. 
Sheep. 
Sheep . 
Dog... 
Dog-... 
Dog-... 
Dog-... 
Dog-... 
Dogr... 
Man3 . . 
Man4.. 
Man5.. 
Man6 . 
Man7.. 


Age 


Older 

Older 

Older 

Older 

Younger 

Younger 

Younger 

Younger 

18  yrs. 


Man 

Child8. 
Child9. 
ChildW. 
Child". 

Cat 

Rabbit. 

Guineapig New-born 

Bird Young 


10  yrs. 

6-7  yrs. 
7-8  yrs. 
2  yrs. 
Foal 
Older 
Older 
Older 

Young 
20  hrs. 
24hrs. 
6  wks. 
8  wks. 
8  wks. 
Older 
Older 
Younger 
Younger 
Older 
Older 
Older 
5  wks. 
Stillborn 

10  wks. 

88  yrs. 

70  yrs. 

70  yrs. 

61  yrs. 

56  yrs. 


34  yrs. 

2  yrs. 

16  mos. 
13.5  mos. 

7  mos. 
New-born 
New-born 


P20fi 


0.1152 
0.2309 
0.2378 
0.2010 
0.2515 
0.3193 
0.3198 
0.5950 
0.0787 
0.0963 
0.1055 
0.1749 
0.1709 
0.3741 
0.1842 
0.2092 
0.2105 
0,1992 
0.4564 
2.6538 
2.7413 
4.1555 
2.5834 
2.4800 
0.1529 
0.2253 
0.3254 
0.5732 
0.3235 
0.1777 
0.3081 
1.6418 
3.3170 
0.8376 
0.1613 
0.2433 
0.2052 
0.1947 
0.1783 

0.2905 

1.1770 

2.1930 

2.5730 

5.3839 

2.919 

2.306 

1.520 

2.25 


Fe203 


0.0243 
0.0266 
0.0283 
0.0223 
0.1105 
0.1085 
0.1254 


0.0254 


0.0379 
0.0975 
0  0750 
0.0486 

6.' 0396 
0.0264 
0.0145 


1.29 
1.02 
0.2960 


0.1460 

0.0138 

0.0191 

0.0249 

0.0839 

0.1672 

0.0542 

0.0489 

0.4437 

4.35! 

0.3214 

0.0354 

0.1157 

0.0668 

0.0693 

0.0807 

0.0552 

1.025 

0.2237 

6.'8049 


Lecithin 


1.31 
2.62 
2.70 
2.28 
2.86 
3.63 
3.64 
6.76 
0.89 
1.09 
1.20 
1.98 
1.94 
4.25 
2.09 
2.38 
2.39 
2.96 
5.18 
30.16 
31.16 
47.23 
29.36 
28.19 
1.73 
2.56 
3.70 
6.51 
3.67 
2.02 
3.50 
18.66 
37.70 
9.52 
1.83 
2.76 
2.33 
2.21 
2.02 

3.30 
13.38 
24.93 
29.24 
61.19 
33.18 
26.21 
17.34 
25.63 


bottle  nursed 
2Badly  nourished 


8Lung  emphysema 
^Stomach,  liver,   kidney  cancer 
5Pleurisy 

6Influenza,  inflammation 
Kidney  inflammation,  enlargement  of  the 
heart 

8Pleurisy 

9Bronchopneumonia 
10Lung  emphysema 
"Pneumonia 


Bolle  (1910a,  1910b)  submits  many  lecithin  estimations  on  the 
marrow  of  several  species  of  animals,  and  says  that  his  findings 
agree  with  those  of  Glikin  in  that  lecithin  seems  to  be  always  pres- 
ent in  the  fat  of  bone  marrow,  the  amount  decreasing  with  advanc- 
ing age,  and  that  in  paralysis  there  is  an  impoverishment  in  this 
particular.     The  tables  did  not  seem  to  the  abstractor  to  show  clear 


120 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


evidence  of  decrease  with  age,  at  least  they  did  not  exhibit  anything1 
like  a  regular  gradation  of  lecithin  according  to  age.  The  method 
of  lecithin  estimation  is  open  to  considerable  improvement. 

Suzuki  and  Yoshimura  (1907)  found  lecithin  in  bones,  and  also, 
an  acid-soluble  organic  phosphorus  compound. 

For  other  lecithin  figures  on  bone  marrow  see  Glikin  1909a,  un- 
der Phosphorus  Metabolism  in  Paralysis. 

Phosphorus  Compounds  in  Cartilage.  Grandis  and  Copello 
(1902)  state  that  the  epiphyseal  cartilage  of  the  femur  and  tibia  of 
the  calf,  compared  with  the  articular  cartilage,  contains  decidedly 
more  phosphorus,  a  part  of  which  is  held  in  organic  combination. 

PHOSPHORUS  COMPOUNDS  OF  MUSCLES 

The  studies  on  the  amounts  of  phosphorus  in  muscles,  and  on 
the  forms  in  which  it  is  present,  have  a  two-fold  significance,  (1)  as 
related  to  the  phosphorus  requirement  of  the  organism,  and  (2)  as 
bearing  on  the  value  of  meats  as  foods.  Muscles  contain  phospho- 
rus in  the  forms  of  nucleoproteins,  phosphatids,  nucleon  (phospho- 
carnic  acid)  and  inorganic  phosphates. 

STUDIES   OF   THE  AMOUNT  AND   DISTRIBUTION   OF  PHOSPHORUS 

Katz's  (1896)  ash  analyses  on  muscle  are  often  quoted  because 
his  was  the  first  extensive  study  of  the  kind,  and  it  covers  many 
species  of  animals.      (See  table  on  following  page.) 

More  recent  studies  are  those  of  Grindley  and  associates,  from 
the  latest  of  which,  by  L.  D.  Hall  and  Emmett  (1912),  we  quote 
the  following : 

PHOSPHORUS  IN  THE  BONELESS  FLESH  OF  DIFFERENT  CUTS  OF 
BEEF  (Hall  and  Emmett,  1912)  Percent 


Soluble   phosphorus 

Insoluble 

Wholesale  cuts 

Inorganic 

Organic 

Total  soluble 

Total 

0.093 
0.094 
0.085 
0.091 
0.085 
0.092 
0.072 
0.062 
0.065 
0.050 
0.048 

0.032 
0.033 
0.024 
0.015 
0.013 
0.016 
0.017 
0.022 
0.019 
0.015 
0.006 

0.125 
0.127 
0.109 
0.106 
0.098 
0.108 
0.089 
0.084 
0.087 
0.065 
0.054 

0.059 
0.046 
0.052 
0.052 
0.050 
0.033 
0.053 
0.051 
0.043 
0.039 
0.023 

0  184 

Clod    

Chuck     

0.173 
0.161 
0  158 

0  148 

0.141 
0  142 

0  135 

Rib      

0.130 
0.104 
0.077 

Plate     

Flank    

See  also  Grindley  and  Emmett  (1905),  Emmett  and  Grindley  (1906,  1909a, 
1909b). 

For  materials  on  the  phosphorus  compounds  of  the  muscle  of  swine,  as 
affected  by  foods,  see  Hart,  McCollum  and  Fuller  (1909)  under  Nutr.  Val.  Org. 
and  Inorg.  P.,  and  Forbes  (1909)  under  Comm.  Foods  in  Rel.  to  P.  Met. 


PHOSPHORUS  METABOLISM 


121 


GO 


p 

o 

K 
O 

w 

CO 

W 

•J 

O 

CO 

H 

H 
< 

w 


a  <u 


t^COt— t  C^  r-H  O -*  T*  CO  C 

romr-^Hiaincoi—cMc 

t~t~r~030ocoo3aoo50o.— icoc 

ooooo'ooooo'i-ho»-! 


iooO'*cocoo~Hiooa3'*tico'* 

NrHIMNrHNCOWHNMOrH 
OOOOOOOOOOt-JoO 


OOOt-Hi— IOHOOOOOH 

oooooooo'ooooo 


05CDt--rtO00C2CCC5C^lO-*lO 
COmrHNr»0)OrHNrHtDlO> 
HHHHHHNHOHOOO 

oooooooo'ooooo 


CD-stHcqcncocoMcorocor—  oio 
c<icD003C<ioo''*'~H~t<cNia3crcco 

laiOmmt-COCOOCDOCtfSCOOO 

oooooooo'ooooo 


CDOOOOOCOeMOOCD-^Ot^-Oira 
0^03CN110CN]r~OCOCO<^a3CO 
■fliXOOJOffi^HHHONW 

OOOOt— tr-lOOOr-(00'— I 


r^-r*ocot-^-^omc~aocDGOo 

r-OOMr- ICMO*— !■— (CM00TFIO 
OHHHr- (i-Ht— (*—(?— (i— I  O  O  t— t 

OOOOOOOOOOOOO 


COOOOOD-OOO^Ht-HCOCMOO'-HO} 

r-- 02  cc  v: :    :':  .*:         :  m  co  cc  <M 

OOOOOOOOOOHHn 
OOOOOOOOOOOOO 


in0005CDCOCOrOC<110CDOOCCD 

NHr- (incMcoost^cijcoo^o 

OOHOOOOOOOOOO 

ooooooooooooo 


COOHOCMOC-sHOCMOOmCOCNICO 

ait^co-rtiOoaDoooo^HOO-* 

CNHOCMCOCM-H'^tlCNieOCOLOOf— t 
OOOOOOOOOOOOO 


OiC0O*#eC0300CDOC0*-iOC0 


COa3005r~COCNI'*IOOCNl-*OCO 

mooaococ4ao-»>— icococoi-ico 

CNJ  CN  «j  >0  lO  CD  CO  lO  OO  iH  O  CO  03 

r-r~r~r^t~t~ir~r~coaooc5or- 


§ftOOQ«OOMfcWBd( 


122 


•OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


A.  Kossel  (1882)  reported  the  following  estimations  of  total 
and  nuclein  phosphorus  in  the  muscle  of  cattle  and  hens. 


TOTAL  AND  NUCLEIN  PHOSPHORUS  IN  MUSCLE 
Kossel  (1882)  Percent 

Species 

H3P04  in 

fresh 
organ 

Nuclein 
HsPOi 

Nuclein 
H3P04   in 
total  H3P04 

Embryonic 

muscle 

Cattle 

Cattle 

Hen, fasted 

' '  well  fed 

0.462 
0.610 
0.838 
0.663 

0.1490 
0.0923 
0.0542 
0.0442 

32.2 

15.1 

6.5 

6.7 

Embryo   28  cm. 

long 

The  distribution  of  phosphorus  in  the  muscle  of  salmon  in  nor- 
mal condition  is  reported  by  D.  Noel  Paton  and  his  associates 
(Paton,  1898;  Paton  et  al.  1897-98)  as  follows: 

DISTRIBUTION  OF  PHOSPHORUS  IN  MUSCLES  OF  SALMON 
Paton,  et  al.  (1897-98)  Percent 


Fish    number 

Type  of  muscle 

Ether    extract 
"Lecithin" 

W  ater  extract. 
"Phosphates" 

Residue,  "Nucleins 

and 

pseudonucleins" 

Total 

14 
14 

76 
76 

Thick 
Thin* 

Thick 
Thin 

0.042 
0.046 
0.060 
0.060 

0.131 
0.094 
0.095 
0.119 

0.056 
0.041 
0.055 
0.063 

0.228 
0.181 
0.210 
0.242 

C1)      The  "thin"  muscle  is  said  to  comprise  about   %    of  the  entire  body. 

Milroy's  (1908)  determinations  on  the  muscle  of  herring  show 
wide  and  irregular  variations  from  month  to  month,  ranging  from 
0.45  to  0.82  percent  P205  in  fresh  muscle. 

Urano  (1907)  has  made  complete  mineral  analyses  of  the  mus- 
cular tissue  of  frogs,  and  also  of  the  press  juice  from  the  same,  the 
blood  plasma,  and  the  muscle  extraction  residue.  The  salts  of  the 
blood  and  lymph  in  the  muscle  were  removed  by  treatment  with 
isotonic  (6  percent)  cane  sugar  solution;  after  which  the  muscle 
juice  was  expressed  by  a  pressure  of  1000  atmospheres.  This  press 
juice  was  dried,  and  extracted  with  hot  water,  and  the  extract  and 
residue  analyzed  separately.  Blood  plasma  was  prepared  by  centri- 
f  uging.      The  phosphorus  figures  are  as  •stated  below : 

PHOSPHORUS  IN  MUSCLE  OF  FROG— Percent  P04 


Sample 

Juice 

Ash 

Muscle 

A 

0.378 
0.633 
0.368 
0.422 
0.311 
0.529 
0.531 
0.109 
0.432 
0.468 

69.399 

64.9 

76.9 

48.0 

63.8 

60.849 

60.236 

16.732 

52.558 

55.469 

0.2268 

B 

0.3861 
0.2245 

0.2532 

C 

0.2084 

0.529 

0.531 

D 

0.109 

0.432 

0.468 

PHOSPHORUS  METABOLISM  123 

Richet  (1900)  analyzed  beef  muscle  serum.  A  kilogram  of  the 
serum  contained  8.9  gm.  ash,  of  which  3.15  gm.  was  P205.  The 
form  in  which  the  phosphorus  was  present  in  the  serum  was  not  de- 
termined. 

Whitfield  (1894)  made  a  special  examination  to  determine 
whether  the  myosin  of  muscle  is  a  nucleoalbumin  and  whether  there 
is  any  nucleoalbumin  in  muscle,  and  decided  that  myosin  is  not  a 
nucleoalbumin  and  that  muscle  contains  no  nucleoalbumin.  Pekel- 
haring  (1896),  however,  isolated  from  the  muscles  of  rabbit,  dog 
and  ox  substances  which  he  identified  as  nucleoprotein. 

Martin  Muller's  (1897)  nucleon  determinations  in  human 
muscle  vary  from  0.1123  to  0.2174  percent,  in  the  muscle  of  adults, 
and  from  0.0000  to  0.0570  in  the  muscle  of  new-born  infants.  Panella 
(1902a,  1903b,  1903e,  1903f,  1904b,  1906b)  decided  that  phospho- 
carnic  acid  is  a  constant,  normal  constituent  of  the  striated  muscles 
of  dogs  and  rabbits,  being  found  in  both  the  red  and  the  white 
muscles  of  rabbits ;  but  his  paper  of  1906  throws  doubt  on  the  quan- 
titative value  of  all  earlier  determinations,  as  they  may  have  been 
made  on  too  small  samples.  The  amounts  reported  at  that  time  are 
0.01  percent  of  fresh  and  0.02-0.03  percent  of  dry  muscle. 

Bajmakov  (1904)  (through  Biochem.  Centralbl.)  reports  an- 
alyses of  the  muscles  of  calves  (5)  and  of  children  (9)  of  different 
ages,  which  show  that  the  dry  substance,  ash,  phosphorus  and  iron 
content  of  muscle,  as  well  as  the  total  protein  and  "organized  pro- 
tein," increase  with  advancing  age.  The  iron  is  said  to  increase  at 
a  more  rapid  rate  than  the  phosphorus.  J 

MacLean  (1912b)  has  made  a  careful  purification  and  examin- 
ation of  the  phosphatids  of  horse  muscle,  and  decides  that  the  chief 
one  present  is  a  lecithin,  the  proportion  of  N:P  being  as  1:1. 

Cavazzani  (1904b)  found  the  phosphocarnic  acid  content  of 
oysters  to  vary  with  the  stages  in  the  life  cycle  from  0.1942  to 
0.5978  percent  with  a  mean  value  of  0.3725. 

COMPARISONS  OF  STRIATED  AND  NON-STRIATED  MUSCLES 

There  seem  to  be  differences  in  chemical  composition  between 
the  striated,  or  voluntary,  and  the  non-striated,  or  involuntary 
muscles;  and  the  heart  muscle  (myocardium),  which  is  striated  but 
involuntary,  differs  somewhat  from  both.  The  reports  as  to  these 
differences  are  not  closely  concordant,  but  we  give  such  as  there  are. 

Saiki  (1908)  was  apparently  the  first  to  make  determinations 
on  the  mineral  constituents  of  non-striated  mammalian  muscle.  He 
used  the  same  analytical  methods  as  Katz.  Saiki  is  alone  in  finding 
the  sodium  content  higher  than  the  potassium  in  non-striated 
muscles,  so  far  as  we  know,  though  others  do  find  somewhat  more 
of  sodium  and  more  of  chlorine  than  in  striated  muscles. 


124  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

MINERAL  ANALYSES  OF  NON-STRIATED  MUSCLES  OF  PIG 

Saiki  (1908) 


Source 

Water 
Percent 

Percent,    dry,    fat-free   substance 

K 

Na 

Fe 

Ca 

Mg 

CI 

S 

P 

Stomach  I 
Stomach  II 
Urinary 

bladder  I 
Urinary 
bladder  II 

81.4 
80.8 

80.6 

79.6 

0.415 
0.332 

0.342 

0.204 

1.549 
1.095 

1.058 

1.045 

0.028 
0.036 

0.042 

0.049 

0.111 
0.117 

0.205 

0.187 

0.026 
0.023 

0.013 

0.012 

0.922 
0.808 

0.816 

0.786 
0.833 

0.444 
0.445 

0.424 

0.373 

0.422 

0.381 
0.428 

0.416 

0.437 

Average 

80.6 

0.323 

1.187 

0.039 

0.155 

0.019 

0.416 

Edw.  B.  Meigs  and  L.  A.  Ryan  (Meigs  and  Ryan,  1912;  Ryan 
and  Meigs,  1912)  report  finding  in  the  water-extract  of  the  smooth 
muscle  of  the  stomach  of  the  frog  (Rana  catesbiana)  0.0958  and 
0.0919  parts  phosphorus  per  100  parts  of  muscle;  in  the  alcohol  ex- 
tract 0.0356  and  0.0260  parts,  and  in  the  residue  0.0146  and  0.0105 
parts,  making  in  the  whole  muscle  0.1460  and  0.1284  parts  phos- 
phorus. They  conclude  that  "1.  The  fibres  of  this  tissue  are  not 
surrounded  by  semi-permeable  membrane.  2.  Most  of  the  water 
of  the  smooth  muscle  fibres  is  held  by  the  colloids  of  the  living  tissue 
as  organic  water.  3.  Most  of  the  potassium,  phosphorus,  sulphur, 
and  magnesium,  which  appear  in  the  ash  of  smooth  muscle,  are  pres- 
ent in  the  living  tissue  in  a  non-diffusible  form."  A  large  portion 
of  the  phosphorus  is  thought  to  be  in  the  form  of  lipoid.  The  fol- 
lowing are  their  ash  analyses  of  striated  and  smooth  muscles  of  the 
frog: 

MINERAL  CONSTITUENTS  OF  SMOOTH  AND  STRIATED  FROG  MUSCLE 
Meigs  and  Ryan  (1912)  Percent,  Fresh  Basis 


K 

Na 

Fe 

Ca 

Mg 

P 

CI 

S 

H20 

0.3483 
0.3518 

0.0572 
0.0500 
0.0536 

0.0099 
0.0094 
0.0096 

0.0007 
0.0007 

0.0335 
0.0228 
0.0281 
0.0042 
0.0042 
0.0042 

0.0288 
0.0313 
0.0300 
0.0132 
0.0126 
0.0129 

0.1554 
0.1541 

0.1547 
0.1460 
0.1284 

0.0650 
0.0674 

0.1492 
0.1322 

79.99 
79.74 

0.3500 
0.3063 
0.3437 
0.3250 

0.0662 

0.1407 

79.87 

0.0648 
0.0804 

0.1191 
0.1200 

0.1724 
0.1501 
0.1612 

82.61 

82.01 

0.0726 

0.0007 

0.1372 

0.1195 

82.30 

According  to  Vincent's  (1902)  comparison  of  the  proteins  of 
smooth  and  striated  muscles,  the  smooth  contain  6-8  times  as  much 
nucleoprotein  as  the  striated,  and  the  heart  muscle  has  an  inter- 
mediate place  in  this  regard.  Panella  (1904b)  found  more  nucleon 
in  non-striated  than  in  striated  muscles.  Balke  and  Ide  (1896) 
found  in  the  heart  of  the  horse  0.116  and  0.105  percent,  and  in  that 
of  the  dog  0.253  percent  of  nucleon. 


PHOSPHORUS  METABOLISM 


125 


Krehl  (1893)  found  that  the  lecithin  content  of  the  healthy- 
heart  muscle  is  about  constant  at  4.2  to  4.6  percent,  while  in  differ- 
ent maladies  it  varies  from  1.1  to  6.3  percent  without  reference  to 
the  state  of  nutrition. 

Rubow  (1905)  studied  the  composition  of  the  heart  and  of  or- 
dinary striated  muscle  as  affected  by  various  conditions.  From  this 
investigation  we  quote  the  following  data  which  show  heart  muscle 
to  be  richer  in  lecithin  than  is  skeletal  striated  muscle. 

LECITHIN  CONTENT  OF  HEART  AND  ORDINARY  STRIATED  MUSCLE 

Percent  of  Muscle 


Subject  and  condition 

Muscle 

No.  of  subjects 

Ether  extract 

Lecithin 

Fat 

Dog,   starved   19-22    days 

Dog,  well  fed 

Dog,   starved   19-22   days 

Heart 

Heart 

Striated 

Striated 

Heart 

Back 

Heart 

Shoulder 

9 
2 

2 
1 

1 
1 
1 

12.23 
11.18 

10.07 
6.07 
18.71 
15.03 
17.62 
14.54 

8.01 
7.49 
5.08 
3.41 
7.54 
2.50 
8.02 
4.15 

4.21 

3.69 

5.99 

2.66 

10.17 

12.53 

9.60 

10.39 

Erlandsen  (1906,  1907)  made  an  exhaustive  study  of  the  phos- 
phatids  of  muscle  from  the  heart  and  from  the  upper  thigh  of  the 
ox,  distinguishing  different  kinds  of  phosphatids.  Both  forms  of 
muscle  contained  lecithin,  a  little  of  a  protagon-  or  jecorin-like  sub- 
stance, a  new  monamino-diphosphatid,  "cuorin,"  and  a  diamino- 
monophosphatid.  Phosphorus  was  also  found  as  phosphocarnic 
acid,  inorganic  salts  and  in  some  nitrogenous  relation  not  identified. 
The  heart  muscle  was  the  richer  in  phosphatids  in  general,  and  es- 
pecially in  the  two  new  phosphatids.  A  high  content  of  cuorin 
seems  to  be  characteristic  of  heart  muscle.  It  contains  less  of  phos- 
phocarnic acid  than  the  thigh  muscle.  The  phosphatids  were  thought 
to  be  present  in  both  types  of  muscle  partly  in  a  free  state  and  part- 
ly combined  with  protein. 

From  the  values  given  in  the  table  below,  Costantino  (1912) 
draws  the  conclusions: 

"1.  There  is  no  measurable  variation  between  striated  and 
non-striated  muscle  in  respect  to  total  phosphorus.  Heart  muscle 
tissue  shows,  however,  a  higher  percentage  of  phosphorus. 

"2.  There  is  an  important  difference  in  respect  to  plain  and 
striated  muscle  content  of  inorganic  phosphorus.  Plain  muscle 
contains  about  the  same  amounts  of  organic  and  inorganic  phospho- 
rus ;  striated  muscle  contains  a  much  higher  percentage  of  inorganic 
than  organic  phosphorus.  Heart  muscle  shows  the  same  relation 
as  plain  muscle. 


126 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


"3.  Both  the  plain  and  heart  muscle  tissue  show  higher  values 
of  organic  than  phosphatid  phosphorus.  Phosphatid  phosphorus 
is  shown  to  be  about  y%  as  much  as  organic  phosphorus  in  plain 


muscle,  and  only  1/3  as  much  in  case 
uterus." 


of    muscle    fibres    of    cow's 


PHOSPHORUS    OF     STRIATED     AND     NON-STRIATED     MUSCLE     OF 
CATTLE  (Costantino,  1912) 


Percents  of  dried  muscle 

Percents  of  total  phosphorus 

Organi- 

Type 

Total 

Inor- 

cally 

Phos- 

Organ- 

Inor- 

Organ- 

Phos- 

Organ- 

phos- 
phorus 

ganic 

com- 

phatid 

ic,  not 

ganic 

phatid 

ic,  not 

phos- 

bined 

phos- 

phos- 

phos- 

phorus 

phos- 

phos- 

phorus 

phos- 

phorus 

phatid 

phorus 

phorus 

phatid 

phorus 

9 

0.7575 

0.6126 

0.1449 

0.1242 

0.0207 

81.50 

18.50 

16.39 

2.11 

? 

0.6498 

0.5287 

0.1211 

0.1059 

0.0152 

81.36 

18.64 

16.29 

2.35 

Heart 

1.0814 

0.4327 

0.6487 



40.01 

59.99 

Heart 

1.0430 

0.3951 

0.6479 

0.4407 

0.2072 

37.88 

62.12 

42.25 

19.87 

Non-striated 

Retractor 

penis 

0.5670 

0.2313 

0.3357 

40.79 

59.21 

Retractor 

penis 

0.5917 

0.2839 

0.3078 

0.1439 

0.1639 

47.97 

52.03 

24.33 

27.70 

Stomach 

0.6010 

0.3148 

0.2862 

0.1518 

0.1344 

52.38 

47.62 

25.25 

21.371 

Stomach 

0.5751 

Uterus 

0.8449 

0.3353 

0.5096 

0.1644 

0.3452 

39.68 

60.32 

19.46 

40.86 

Uterus 

0.7329 

0.2142 

0.5187 

0.2081 

0.3106 

29.22 

70.78 

28.39 

42.39 

(x)      The  compilers  compute  this  to  be  22.36. 

MacLean  (1913)  found  heart  muscle  to  contain  lecithin,  cuorin 
and  a  trace  of  diamino-monophosphatid  which  resembles  the  sub- 
stance described  by  Stern  and  Thierf  elder  and  by  Thudichum,  An 
analogous  compound  has  been  obtained  from  kidneys  by  Dunham 
and  Jacobson  and  by  MacLean. 

For  the  distribution  of  phosphorus,  potassium  and  chlorine 
within  the  muscle  fiber  see  Menten  (1909). 


PHOSPHORUS  IN  FEATHERS 

In  connection  with  Weiske's  study  (1889)  of  the  effects  of  age 
on  the  composition  of  the  bodies  of  birds,  analyses  were  made  of  the 
feathers.  From  the  following  data  it  appears  that  as  a  growing 
fowl  increases  in  age  the  phosphorus  of  the  feathers  decreases  in 
percentage  during  the  first  18  weeks.  At  the  18th  week  the  feath- 
ers contain  a  lower  percentage  of  mineral  matter  generally  than 
at  any  other  time  during  the  first  year,  and  this  is  also  the  low  point 
for  percentage  content  of  calcium,  sulphur  and  phosphorus. 


PHOSPHORUS  METABOLISM 

ANALYSES  OF  FEATHERS  OF  BIRDS 
Weiske  (1889)  Percent  of  Dry,  Fat-free  Feathers 


127 


Bird 

Age, 
■weeks 

Mineral 
matter 

CaO 

MgO 

S02 

P205 

Feed 

0 

2.04 

0.31 

0.140 

0.11 

0.190 

Corn 

1 

1.98 

0.16 

0.051 

0.23 

0.190 

Corn 

2 

2.19 

0.45 

0.047 

0.48 

0.160 

Corn 

4 

2.09 

0.40 

0.120 

0.27 

0.150 

Corn 

8 

1.51 

0.15 

0.044 

0.25 

0.140 

Corn 

12 

1.50 

0.19 

0.050 

0.14 

0.140 

Corn 

18 

0.88 

0.15 

0.022 

0.052 

0.060 

Corn 

24 

1.80 

0.40 

0.049 

0.059 

0.117 

Corn 

34 

1.78 

0.62 

0.024 

0.10 

0.075 

Corn 

44 

2.11 

0.87 

0.011 

0.22 

0.067 

Corn 

52 

1.82 

0.82 

0.031 

0.15 

0.062 

Corn 

52 

1.89 

0.80 

0.041 

0.13 

0.070 

Corn 

0.40 

0.050 

0.016 

0.037 

0.074 

Meat 

0.62 

0.092 

0.006 

0.147 

0.133 

Meat 

THE  PHOSPHORUS  OF  BRAIN,  NERVES  AND  CEREBROSPINAL  FLUID 

HISTORICAL  REVIEW  TO  THE  TIME   OF  THUDICHUM 

None  of  the  general  analyses  of  brain  before  those  of  Thudi- 
chum need  receive  especial  attention.  Thudichum  mentions  one 
by  Hensing  (1719)  when  the  phosphorus  of  the  brain  was  dis- 
covered. Fourcroy's  (1793)  work  was  quite  extensive  and  Vau- 
quelin's  (1811)  was  significant  enough  to  have  led  to  its  translation 
from  the  French  into  both  English  and  German,  and  to  be  reviewed 
at  some  length  in  Thudichum's  "Die  chemische  Konstitution  des 
Gehirns."  Couerbe's  (1834)  analyses  were  followed  by  Fremy 
(1841,  1842).  We  find  early  ash  analyses  by  Breed  (1851b)  and 
by  Geoghegan  (1877-78).  VonBibra  (1853, 1854a)  made  phosphorus 
determinations  on  the  fat  obtained  from  the  brains  of  fifteen  kinds 
of  animals,  and  of  men,  and  showed  that  this  phosphorus  is  a  con- 
stituent of  the  fat  itself.  The  next  year  (vonBibra,  1854b)  he 
reports  phosphorus  in  the  spinal  cord  and  nerves  of  several 
species,  and  concludes  that  in  brain  and  spinal  marrow  the  phospho- 
rus content  depends  on  the  fats  containing  phosphorus,  and  in 
nerves  principally  on  the  cerebric  acid.  Liebreich's  (1865)  analysis 
is  significant  for  its  discovery  of  protagon,  which,  however,  is  now 
known  not  to  be  a  single  compound.  After  Gobley  and  others  made 
the  various  studies  of  "lecithin,"  Gobley  (1877)  reported  a  general 
analysis  of  brain.  The  later  one  of  Gutnikov  (1896-97)  should  also 
be  mentioned. 

Some  of  these  old  reports  make  interesting  reading.  Couerbe 
(1834),  finding  a  lower  phosphorus  content  in  the  fat  of  an  idiot's 
brain  and  a  higher  in  that  of  the  insane  than  normal,  concludes: 
"Phosphorus  is  the  exciting  principle  of  the  nervous  system,"  and 

Note:  For  general  discussions  of  the  subject  of  the  chemical  composition  of  the  brain 
and  nerves  the  reader  is  referred  to  Thudichum  (1901),  Halliburton  (1901b,  1905>  Coriat 
(1905)    and   Frankel    (1909a). 


128  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

"Absence  of  phosphorus  from  the  brain  reduces  man  to  the  level  of 
the  brute,  while  a  great  excess  irritates  the  nervous  system,  caus- 
ing mental  alienation." 

THE  WOEK  OF  THUDICHUM 

At  the  time  of  his  report  in  1875  Thudichum  (1875)  had  anal- 
yzed a  thousand  brains,  mostly  normal  brains  from  human  subjects, 
and  had  found  and  identified  many  of  the  compounds  which  he  af- 
terwards described  and  classified  more  fully.  "Die  chemische  Kon- 
stitution  des  Gehirns"  appeared  in  1901  and  seems  to  have  greatly 
stimulated  and  influenced  later  work  on  brain  chemistry  in  general 
and  especially  on  the  chemistry  of  lipoids.  The  classification  of 
compounds  found  is  given  below;  also  analytical  determinations  of 
the  principal  constituents  of  the  white  and  the  gray  matter : 

THUDICHUM'S  CLASSIFICATION  OF  THE  BRAIN  CONSTITUENTS 

A.     GEOUP   OF   PHOSPHOEUS-CONTAINING   CONSTITUENTS    OE   PHOSPHATIDS 
Subgroup  of  the  monophosphatids  containing  one  nitrogen.  N:P— 1:1. 

Lecithin 

Cephalin 

Paramyelin 

Myelin 
Subgroup  of  monophosphatids  containing  two  nitrogens.  N:P=2:1 

Amidomyelin 

Amidocephalin 

Sphingomyelin 

Apomyelin 
Subgroup  of  diphosphatids  containing  two  nitrogens.      N:P=2:2. 

Assurin 
Subgroup  of  nitrogen-containing  phosphatid-sulphatids 
Subgroup  of  nitrogen-free  monophosphatids 

Lipophosphoric  acid 

Butophosphoric  acid 

B.  GEOUP    OF   NITEOGEN-CONTAINING   PHOSPHOEUS-FEEE    CONSTITUENTS      ' 

Subgroup  of  cerebrosids  or  cerebrogalactosids 

Phrenosin 

Kerasin 
Subgroup  of  cerebrin  acids 

Cerebrinic  acid 

Sphaerocerebrin 
Subgroup  of  cerebrosulphatids 
Subgroup  of  amidolipotids  or  nitrogen-containing  fats 

Bregenin 

Krinosin 
Subgroup  of  alkaloids 

Hypoxanthin 

Gladiolin 

Tenysin 
Subgroup  of  amino  acids  and  amids 

Leucin  and  its  homologues 

Tyrosin 

Urea 

C.  GEOUP  OF  CONSTITUENTS  WHICH  CONTAIN  ONLY  THEEE  ELEMENTS 

Subgroup  of  nitrogen-free  alcohols 

Cholesterin 

Phrenosterin  (?) 
Subgroup  of  carbohydrates 

Inosite 

Glycogen  (?) 


PHOSPHORUS  METABOLISM 


129 


Subgroup"  of  nitrogen-free  organic  acids 
Formic  acid 
Sarcolactic  acid 
Succinic  acid 
Oxyglyceric  acid  (?) 
D      GROUP  OF  ORGANOPLASTY  OR  ALBUMINOUS  SUBSTANCES 

Subgroup  of  nitrogen-containing  sulphatid-phosphatids 

Neuroplastin 

Gangliocytin,  cytophosphatid-  or  nuclein-substances 
Subgroup  of  nitrogen-containing  sulphatids 

Albumen 

Collagen 

E.  GROUP  OF  INORGANIC  PRINCIPLES,  ACIDS,  BASES  AND  SALTS 

These  are  found  in  part  in  the  water- extract,  in  part  in  combination 
with  many  of  the  before-named  constituents. 
Sulphuric  acid 

Hydrochloric  acid  and  chlorine  in  chlorides 
Phosphoric  acid 
Carbonic  acid 


Potassium 

Sodium 

Ammonium 

Calcium 

Magnesium 

Copper 

Iron 

Manganese 

Aluminum,  silicic  acid,  fluorine 

SUMMARY  OF  ANALYSES  OF  BRAIN 


In  compounds  forming  bases  with  products 
obtained;  or  in  compounds  with  phosphoric 
acid,  and  then  combined,  as  phosphates,  with 
these  products;  or  in  combination  with  min- 
eral acids  as  free  mineral  salts  in  the  fluids 
and  extracts. 


Gray  matter 
Percent 

White  matter 
Percent 

Ether-extract  with  cephalins,   lecithins 

85.270 
7.608 

1.950 
0.424 
0.500 
0.193 
0.102 
0.017 
0.025 
0.092 

70.230 
8.630 

11  497 

Cerebrosids,  cerebrin  acids  and  myelin.  . 

6.910 
1.403 

0.2171 
0.0456 

0.1717 

OTHER  STUDIES  OF  THE  COMPOUNDS  PRESENT  IN  BRAIN 

Waldemar  Koch  and  William  H.  Goodson  (1906)  give  analyses 
of  two  fractions  of  the  gray  matter,  the  prefrontal  and  the  motor 
areas,  which  do  not  differ  materially  in  the  normal  brain,  and  of  the 
white  medullated  matter  of  the  corpus  callosum,  which  differs  some- 
what from  them,  particularly  in  the  amount  of  cerebrins.  The 
sciatic  nerve  differs  from  the  corpus  callosum  in  having  considerably 
more  protein  and  less  water,  which  may  be  due  to  a  greater  amount 
of  connective  tissue.  The  table  below  gives  the  analyses  together 
with  some  for  the  study  of  the  effects  of  degeneration  of  brain  and 
nerve  tissue.  W.  Koch's  (1904)  analysis  of  epileptic  brain  is  given 
in  the  chapter  on  metabolism  in  disease. 


130 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BTJL.  5 


ANALYSES  OF    NORMAL  AND   DEGENERATED   BRAIN  AND   NERVE 
TISSUE  (Koch  and  Goodson,  1906)  Percent 


Brain  (Human) 

Sciatic  . 

nerve 

(Human) 

Prefrontal  area 

Motor  area 

Corpus 
callo- 
sum 

Spinal  cord 
(Dog) 

Normal 

Degener- 
ated 

Normal 

Degener- 
ated 

Normal 

Degener- 
ated 

Total  solids 

17.5 

15.2 

18.4 

17.4 

30.0 

35.8 

31.5 

28.4 

Percent  of  solids 


Simple  proteids 

Nucleoproteids 

Lecithins 

Cephalins 

Cerebrins 

Sulphur  compound.. 

Extractives  

Inorganic  salts 


25.8 

23.8 

14.8 

8.9 

7.1 

5.9 

11.0 

7.0 


19.1 
37.7 
12.3 
8.5 
7.7 
5.6 
10.8 
6.6 


26.6 

21.0 

12.4 

11.0 

8.6 

5.9 

10.6 

6.0 


12.1 

35.3 
15.0 
15.0 
12.0 

6.0 
10.0 

6.0 


16.7 

11.4 

14.5 

7.6 

17.7 

7.3 

5.8 

2.7 


12.6 
34.9 
7.1 
7.8 
7.2 
10.0 
5.0 
3.6 


18.1 
6.7 

26.4 

15.9 
5.3 
3.7 
2.2 


14.9 
12.8 

26.3 

15.1 
6.4 
3.4 
2.0 


Other  single  determinations  of  phosphorus  compounds  reported 
by  different  workers  are  collected  in  the  following  table : 

SOME  DETERMINATIONS  OF  BRAIN  OR  NERVE  CONSTITUENTS 


Compound  and  source 

Author 

Date 

Moist  substance 
Percent 

Dry  substance 
Percent 

Phosphocarnic  acid 

Panella 

1903g  . 
1903h 

0.2050 

0.9037 

Panella 
Panella 
Panella 

1903h 
1903h 
1903h 

0.2142 

0.2245 
0.2837 

1.1803 

1.0044 

1.3273 

Panella 

1903h 

0.3259 

Brain,  calf,  gray  matter 

Panella 

1903h 

0.1917 

Panella 

1903h 

1903h 

0.2871 
0.3502 

1.1841 

1.6937 

Panella 

1903h 

0.3520 

1.5820 

Panella 

1903h 

0.3316 

1.7240 

Lecithin 

Chevalier 

1886 

14.80 

Embryo,  62  cm.  (cattle) 

Raske 

1886 

0.610 

6.6331 

Embryo,  68  cm.   (cattle)    

Raske 

1886 

0.315 

3.4923 

Petrow- 
sky 

1873 

3.1714 

17.2402 

White   matter   of    brain 

Petrow- 
sky 

1873 

3.1347 

9.9045 

Burow 

1900 

3.954 

Nuclein 

Geog- 
hegan  1 

1877-78 

0.1390 

0.1624 
0.1340 
0.1368 

1  Percents  computed  by  compilers. 

Special  studies  of  cephalin  are  reported  by  Thudichum  and 
Kingzett  (1876),  W.Koch  (1902b,  1903),  Cousin  (1906),  Falk 
(1908),  and  by  Frankel,  Neubauer  and  Dimitz  (Frankel  and  Neu- 
bauer,  1909 ;  Frankel  and  Dimitz,  1909) .  Falk  says  that  the  ceph- 
alin of  the  peripheral  nerves  is  not  the  same  as  that  of  brain. 


PHOSPHORUS  METABOLISM 


131 


He  reports  on  the  chemical  composition  of  peripheral  nerves. 
Frankel  (1908,  1909b)  describes  several  brain  lipoids,  and  Argiris 
(1908)  reports  quantitative  estimations  of  phosphorus  compounds 
on  brains  of  birds  and  fish. 

Scott  (1899)  discusses  the  Nissl  granules  of  nerve  cells,  which 
are  of  nucleoprotein  nature  and  are  said  to  consist  of  chromatin 
which  has  diffused  from  the  nucleus  into  the  cytoplasm.  Levene 
(1903c)  gives  further  discussion  of  these  chromatins,  saying  that 
they  are  compounds  of  nucleic  acid  with  one  or  more  proteins,  or 
with  protein  and  carbohydrate.  He  reviews  the  work  on  the  sub- 
ject, considering  the  question  of  the  identity  or  difference  of  nucleic 
acids  from  different  sources.  The  nucleoprotein  which  Levene 
(1899)  isolated  from  brain  and  analyzed  was  said  to  differ  from 
other  nucleoproteins  in  its  low  phosphorus  content,  and  in  having 
a  considerably  higher  amount  of  proteins  bound  to  its  nuclein.  Halli- 
burton's (1894)  paper  on  the  proteins  of  brain  we  have  not  seen. 

The  so-called  "protagon,"  which  was  once  looked  upon  as  the 
most  distinctive  component  of  brain  tissue,  has  been  proved  to  be  a 
mixture,  and  we  have  given  the  history  of  its  study  in  our  chapter 
on  the  chemistry  of  phosphatids.  Noll  (1899)  made  a  number  of 
determinations  of  protagon  in  medullary  tissues,  and  all  of  the  early 
analyses  include  it  as  an  item.  Rosenheim  and  Tebb  (1909c)  find 
that  sphingomyelin,  containing  4  percent  phosphorus,  is  the  most 
important  component  of  the  so-called  protagon'. 

W.  Koch  (1907b,  1910a)  thought  he  had  evidence  of  a  combin- 
ation which  might  be  called  phosphatid-cerebrosid-sulphatid.  Bar- 
bieri  (1909)  describes  cerebroin,  containing  0.7  percent  phosphorus, 
found  chiefly  in  the  gray  matter  of  the  brain  (ox).  It  yielded 
neither  glycerin  nor  sugar.  Bethe  (1902)  reports  a  cerebrinic  phos- 
phoric acid  of  about  1  percent  phosphorus  content  found  in  the 
brain  of  the  horse.  Jolly  (1898)  looked  for  evidence  of  phosphorus 
in  the  organic  portions  of  brain  in  some  other  form  than  such  as 
yields  orthophosphoric  acid  on  simple  ashing,  but  did  not  find  it. 

Masuda  (1910)  reports  total  phosphorus  figures  for  the  three 
divisions  of  the  brain  as  in  the  following  table : 

TOTAL  PHOSPHORUS  CONTENT  OF  THE  THREE  PARTS  OF  THE  BRAIN 
Masuda  (1910)   Percent  P 


Cerebrum 

Cerebellum 

Mid-brain 

0.1838 
0.3324 
0.2545 
0.2813    . 
0.2743 

0.1593 

0.2829 
0.2760 
0.2971 
0.2824 

0.2638 

0.3437 

0.4002 

0.3736 

0.3480 

132 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


For  total  phosphorus  determinations  on  the  brain  of  horse  see 
Baumstark  (1885)  and  Chittenden  (1897),  pig, — Hart,  McCollum 
and  Fuller  (1909),  sheep, — Kutanin  (1910),  and  pigeon, — Funk 
(1912a). 

For  partial  separations  of  the  groups  of  phosphorus  compounds 
in  the  brain  of  the  horse  see  Baumstark  (1885),  and  the  same  for 
sheep,  Chittenden  (1897). 

DIFFERENTIAL    ESTIMATIONS    OF    PHOSPHORUS    COMPOUNDS    OF    BRAIN    UNDER 

SPECIAL  CONDITIONS 

In  their  report  of  "A  Chemical  Study  of  the  Brain  in  Healthy 
and  Diseased  Conditions,  with  Especial  Reference  to  Dementia 
Praecox,"  W.  Koch  and  Mann  (1909)  give  the  following  as  the  parti- 
tion of  phosphorus  among  the  various  classes  of  compounds.  We 
have  brought  together  all  of  those  which  are  spoken  of  as  normal 
and  grouped  the  pathological  cases  as  do  the  authors.  The  values 
are  percents  of  the  total  phosphorus. 

PARTITION  OF  PHOSPHORUS  AMONG  THE  BRAIN  COMPOUNDS 
Koch  and  Mann  (1909) 


6-weeks  child1 

2-yr.  child,  cortex 

2-yr.  child,  corpus  callosum 

2-yr.  child,  whole  by  computation . 

12  yr.  child,  cortex 

12-yr.  child,  corpus  callosum 

12-yr.  child,  whole  by  computation  . 

16-yr.,  Case 20 

24-yr.,Casel9 

43-yr.,  Case  70 

49-yr.,  Case  18 

Dementia  praecox,  Case  28 

Dementia  praecox,  Case  29 

Dementia  praecox,  Case  41 

Dementia  praecox,  Case  42 

General  paralysis,  Case  22 

General  paralysis,  Case  23 

General  paralysis,  Case  24 

General  paralysis,  Case 40 

Melancholia,  Case  25 

Melancholia,  Case  26 

Brain  of  a  dog • 


Total  P 

Percent  of 

solids 


Part 


1.50 
1.46 


1.45 
1.45 


Whole 
brain 

1.72 

1.48 

1.45 
1*50 


1.50 


Percent  of  total  P 


Protein  P 


Part 


Whole 
brain 

5 


4.7 
3.7 
6 
3.9 


3.8 
5.1 
3.6 
4.7 
5.9 
4.8 
4.6 
4.6 
4.2 
4.1 


4.5 


Lipoid  P 


Part 


Whole 
brain 

54 


67 


72 

70.3 

73.1 

75 

71.8 


73.7 
73.0 
73.6 
70.8 
69.3 
70.6 
68.2 
69.9 
69.2 
74.7 


70. 


Extractive  P 


Part 


22 


Whole 
brain 

41 
27 


23 

25.1 

23.2 

19 

24.2 


22.6 
21.9 
22.9 
24.4 
24.7 
24.7 
26.9 
25.6 
26.6 
21.2 


25.5 


(!)      Birth  premature;    brain   underdeveloped. 

The  authors'  conclusions  with  regard  to  the  phosphorus  are : — 
1.  With  growth  of  the  brain  there  is  an  increase  in  the  lipoid  phos- 
phorus and  a  decrease  in  the  extractive  phosphorus.  2.  The 
phosphorus  agreement  between  the  brain  of  man  and  dog  is  quite 
close.      3.     Comparison  of  brains  from  cases  in  which  the  causes  of 


PHOSPHORUS  METABOLISM 


133 


death  were  of  an  entirely  different  character  showed  no  variations 
of  importance.-  4.  The  brains  from  cases  of  dementia  praecox 
showed  no  marked  change  in  the  amount  and  distribution  of  phos- 
phorus as  compared  with  the  normal.  5.  The  results  from  brains 
of  cases  of  general  paralysis  show  that  the  destructive  changes  in 
this  disease  affect  the  brain  generally  and  not  one  constituent  in 
particular.  There  is,  however,  a  tendency  for  the  lipoid  phosphorus 
to  be  decreased,  indicating  a  greater  destruction  of  the  phosphatids. 

CHANGES  OF  THE  BRAIN  WITH  GROWTH 

Koch  and  Mann  (1907)  give  the  following  comparison  of  the 
chemical  composition  of  three  human  brains  at  different  ages,  and 
draw  conclusions  as  quoted : 

COMPARISON  OF  BRAINS  OF  DIFFERENT  AGES— Percents  of  Solids 


6- weeks 

child 

Whole 

2-years  old 

19-year s  old 

Gray 

White 

Whole 

.  Gray 

White 

Whole 

46.6 

48.4 

31.9 

40.1 

47.1 

27.1 

37.1 

12.0 

10.0 

5.9 

8.0 

9.5 

3.9 

6.7 

Ash 

8.3 

5.8 

3.2 

4.5 

5.9 

2.4 

4.1 

Lecithins  and  cepha- 

24.2 

24.7 

26.3 

25.5 

23.7 

31.0 

27.3 

6.9 

8.6 

17.2 

12.9 

8.8 

16.6 

12.7 

Lipoid   S  as   SO* .  . . 

0.1 

0.1 

0.5 

0.3 

0.1 

0.5 

0.3 

Cholesterin    (by  dif- 

1.9 

2.4 

15.0 

8.7 

4.9 

18.5 

11.7 

88.78 

84.49 

76.45 

80.47 

83.17 

69.67 

76.42 

Total    sulphur     .... 

0.52 

0.53 

0.63 

0.58 

0.46 

0.50 

0.48 

Total  P 

1.72 

1.50 

1.64 

1.48 

1.45 

1.45 

1.45 

DISTRIBUTION  OF  SULPHUR— Percent  of  Total  Sulphur 


Protein  S  . 
Lipoid  S  .  . 
Neutral  S  . 
Inorganic  S 


62 

63 

55 

59 

73 

51 

6 

6 

27 

17 

7 

36 

26 

22 

13 

17 

12 

6 

6 

9 

5 

7 

8 

7 

62 

22 

9 

7 


DISTRIBUTION  OF  PHOSPHORUS— Percent  of  Total  Phosphorus 


Protein  P    

Lipoid  P    

Water-soluble  P 


5 

6 

6 

6 

5 

5 

54 

62 

72 

67 

63 

81 

41 

32 

22 

.  27 

32 

15 

5 

72 
23 


"In  the  above  table  is  to  be  observed  with  the  growth  of  the 
brain : 

"A  decrease  in  moisture,  proteins,  extractives,  and  ash,  a 
change  usually  found  in  growing  tissues. 

"An  increase  in  cerebrins,  lipoid  sulphur  and  cholesterin,  in 
other  words  the  substances  which  predominate  in  the  white  matter. 

"The  relative  increase  in  the  lecithin  and  cephalin  is  not  so  ap- 
parent, as  at  an  early  age  the  brain  is  supplied  with  practically  the 
same  proportion  of  lecithin  and  cephalin  as  the  adult ;  in  fact,  it  is 
the  richest  tissue  in  these  constituents,  the  adrenal  coming  next 
with  12  percent  and  the  liver  with  10  percent. 


134 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


"In  the  distribution  of  phosphorus  and  sulphur  there  is  to  be 
observed  a  tendency  of  the  water-soluble  forms  to  become  converted 
into  more  complex  water-insoluble  forms,  principally  lipoids,  which 
increase  much  more  in  actual  amount  than  the  above  increase  in  rel- 
ative amount  might  indicate.  The  variation  in  both  the  protein 
sulphur  and  phosphorus  is  less  than  might  be  expected." 

M.  Cohn  (1907)  gives  the  following  determinations  on  the 
brains  of  children.  No  changes  were  noted  as  characteristic  of  the 
diseases. 

ANALYSES  OF  THE  BRAINS  OF  CHILDREN 
M.  Cohn  (1907) 


Age 

Sex 

Diagnosis 

Wt.  of 
brain 

H2O 

Percent  of  dry  substance 

No. 

Grams 

Percent 

N 

P 

Ca 

1 

Fetus,  25  cm. 

56 

91.1 

9.44 

0.0631 

2 

1  day 

495 

89.30 

9.69 

1.69 

0.0519 

3 

VA  mos. 

M 

Intestinal  catarrh 

402 

87.80 

9.47 

1.64 

4 

3K  mos. 

F 

Naval  sepsis 

547 

86.17 

9.76 

1.58 

0.0231 

5 

7  mos. 

F 

Pneumonia 

656 

83.65 

8.95 

1.68 

0.0285 

6 

8  mos. 

M 

Post  pneumonia  empyema 

.   824 

83.35 

9.09 

1.58 

0.0263 

7 

11  mos. 

F 

Pneumonia,  measles 

846 

83.93 

8.92 

1.56 

8 

VA  yrs. 

F 

Pneumonia,  measles 

801 

82.34 

9.35 

1.62 

0.0237 

9 

2H  yrs. 

F 

Pneumonia,  measles 

994 

82.15 

8.58 

1.55 

0.0197 

10 

Zte  yrs. 

M 

Scarlatina 

1247 

81.57 

8.05 

1.51 

11 

4  yrs. 

M 

Diphtheria 

1304 

80.33 

7.97 

1.50 

0.0191 

12 

6K  yrs. 

F 

Scarlatina 

1280 

80.01 

8.15 

1.51 

0.0181 

13 

20  yrs. 

F 

Erysipelas,  nephritis 

1270 

77.52 

7.57 

1.50 

0.0169 

14 

6%  mos. 

M 

Tetanus 

822? 

84.40 

9.72 

1.71 

0.0282 

15 

10  M  mos. 

M 

Atrophy,  tetanus 

750 

84.31 

9.15 

1.72 

0.0225 

Dhere  and  Maurice  (1909)  made  analyses  of  the  peripheral 
nerves  of  19  dogs  betwen  the  ages  of  4  weeks  and  8  years,  from 
which  they  report  the  following  results,  and  conclude  that  the  phos- 
phorus content  of  nerves  (on  the  dry  basis)  diminishes  with  in- 
crease of  age,  but  the  diminution  does  not  occur  equally  among  the 
different  groups  of  phosphorus  compounds. 

ANALYSES  OF  PERIPHERAL  NERVES  OF  DOGS  OF  DIFFERENT  AGES 

Dhere  and  Maurice  (1909) 


Ages 

No. 

Percents  of  dry  substance 

Percents  of  total  P 

Group 

Total 
P 

Lipoid 
P 

Nuclein 
P 

Inorg'anic 
P 

Lipoid 
P 

Nuclein 
P      . 

Inorganic 
P 

I 
II 

III 

4  wks. — 4  mos. 
6 — 13  mos. 

2-8  yrs. 

9 

6 

4 

0.990 
0.717 

0.605 

0.430 
0.335 

0.285 

0.072 
0.061 

0.056 

0.488 
0.321 

0.264 

43.43 

(     46.72 
-,  (gain  of 
1     7.6/o) 
47.11 
\  (gain  of 
\    8.5*) 

7  27 
I     8.51 

<  (gain  of 
(  17.0*) 

9.26 

<  (gain  0 
I  27.  4*) 

49.30 
I   44.77 
i  (loss  of 
1    9.2*) 

43.63 
\  (loss  of 
I    11.5*) 

See  also  Kutanin   (1910) 


PHOSPHORUS  METABOLISM 


135 


J.  Smith  and  W.  Mair  .(1912)  give  the  following  report  of  a 
study  of  the  lipoid  content  of  the  brains  of  dogs  of  different  ages, — 
a  Utter  of  eight  pups  and  their  mother.  They  find  that,  while  the  to- 
tal lipoids  and  the  cerebrosids  show  a  marked  and  constant  increase 
with  age,  the  percentage  of  phosphorus  and  phosphatids  show  a 
maximum  at  3  weeks,  and  thereafter  a  continuous  decrease,  al- 
though the  absolute  amount  present  increases  constantly.  Atten- 
tion is  called  to  the  fact  that  while  the  milk  contains  only  minute 
traces  of  these  lipoids,  in  the  brain  alone  at  the  age  of  3  to  6  weeks 
there  is  a  daily  addition  of  .045  gm.  phosphatid,  .007  gm.  cerebrosid, 
and  .015  gm.  cholesterin.  This  is  taken  as  evidence  of  a  synthesis 
of  the  compounds  in  the  body. 


LIPOID  CONTENT  OF  DOGS'  BRAINS  AT  DIFFERENT  AGES 
Smith  and  Mair  (1912) 

Age  and  subject 

3  days, 
3  dogs 

3  weeks, 
3  dogrs 

6  weeks, 
1  dog 

12  weeks, 
1  dog 

Adult, 
1  dog 

Human 
adult 

1.38 
23 

5.40 
25 

8.90 
34 

15.0 
38 

19.25 
47 

Percentage  Composition  of  Chloroform  Extract 


Cholesterin  . 
Cerebrosid  .  . 
Phosphorus  . 
Phosphatid  . 
Other   lipoids 


18.3 

16.8 

18.5 

17.1 

21.8 

1.5 

3.5 

6.8 

11.5 

21.4 

2.10 

2.42 

2.27 

2.11 

1.76 

52. 

60. 

57. 

52. 

44. 

28. 

20. 

28. 

19. 

13. 

21.8 
10.7 

1.58 
39.5 
28. 


Amounts  Found  in  the  Brain — Grams 


Phosphatid  . 
Cerebrosid  .  . 
Cholesterin  . 
Other  lipoids 
Total  extract 


0.16 

0.80 

1.70 

2.90 

4.0 

0.005 

0.046 

0.20 

0.67 

1.95 

0.058 

0.22 

0.55 

1.00 

2.0 

0.10 

0.28 

0.55 

1.20 

1.17 

0.32 

1.34 

3.00 

5.77 

9.12 

Messing  (1912)  studied  the  mineral  constituents  of  normal  and 
pathological  brains.  The  P205  content  increases  with  age  and  the 
weight  of  the  brain,  until  about  the  60th  year,  after  which  it  de- 
creases. Arteriosclerosis  causes  a  decrease  of  the  phosphorus,  and 
an  increase  of  both  calcium  and  S03. 

Mathilde  Koch  (1913)  has  made  similar  determinations  to  those 
of  Koch  and  Mann  on  brains  from  pig  fetuses  of  50, 100  and  200  mm. 
length,  and  from  albino  rats  at  birth,  and  in  the  adult  state,  carrying 
on  work  started  by  Waldemar  Koch.  She  finds  that  chemically, 
anatomically  and  physiologically,  the  brain  of  the  albino  rat  is  at 
about  the  same  stage  at  birth  that  the  pig  fetus  is  at  50  or  100  mm., 
and  represents  as  young  nervous  material  as  can  conveniently  be  an- 
alyzed at  present.  The  phosphatids  there  comprise  15.2  percent  of 
the  solids,  and  the  total  phosphorus  1.87  percent.  Of  the  phos- 
phorus 13.3  percent  is  in  protein,  33.1  percent  in  lipoid  and  53.55 


136 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


percent  in  water-soluble  form.  From  examinations  of  rat  brains 
at  1,  10,  20,  40,  120  and  210  days  (Koch,  W.,  and  M.  L.,  1913)  it  is 
learned,  among  other  things,  that  in  the  period  of  the  most  rapid 
growth  and  of  the  beginning  of  medullation,  from  the  10th  to  the 
20th  day,  there  is  most  rapid  development  of  phosphatids.  Phos- 
phatids  are  found  in  both  cells  and  sheaths. 

PHOSPHORUS  IN  THE  LIVER 

Phosphorus  Content.  Szymkiewicz  (1891)  made  sulphur  and 
phosphorus  determinations  on  the  ash  of  some  80  samples  of  the 
livers  of  cattle  of  various  ages,  from  the  13-cm.  fetus  to  the  cow  of 
13  years.  In  the  cases  of  the  smaller  fetuses  a  mixture  of  2-6 
livers  was  used.  There  were  no  characteristic  variations  in  the 
phosphorus  content  of  the  livers  of  the  fetuses  (minimum,  1 .52  and 
maximum,  1.98  percent),  though  there  were  irregular  individual 
variations.  As  a  rule  the  phosphorus  content  after  birth  is  ap- 
preciably lower  than  before  birth,  although  among  the  24  there  were 
some  individual  cases  in  which  the  phosphorus  content  is  higher 
than  the  lowest  found  in  the  fetus.  Among  the  24  there  is  no  evi- 
dence of  any  characteristic  effect  due  to  advancing  age  or  to  sex. 
The  mean  of  the  values  on  calves,  oxen  and  cows  is  1.39  percent  P. 

Kriiger  (1895)  obtained  results  practically  like  those  of 
Szymkiewicz  except  that  his  averages  for  adult  animals  are  some- 
what lower  than  for  calves.  In  the  livers  from  human  subjects 
the  phosphorus  was  considerably  higher  in  the  new-born  infants 
than  in  the  adults. 

AVERAGE  PHOSPHORUS   (P)   CONTENT  OF  LIVERS  OF  CATTLE  OF 
DIFFERENT  AGES  (Friedrich  Kriiger,  1895)   Percent  Dry  Basis 


Fetuses,  length  in  centimeters 

Calves 

Cows 

Oxen 

30-40 
1.74 

40-50 
1.73 

50-60 
1.73 

60-70 
1,65 

70-80 
1.69 

/ 

80-90    90-100 
1.72 

1.46 

1.29 

1.30 

Weiske's  (1886)  experiment  testing  the  effects  on  lambs  of 
feeding  hay  which  had  been  treated  with  dilute  sulphuric  acid 
showed  the  phosphorus  content  of  the  livers  of  the  3  lambs  on  such 
feed  and  on  normal  feed  to  range  between  0.450  and  0.  485  percent 
P,  with  the  mean  value  of  0.468  percent. 

Paton  (1895-6),  in  discussing  the  relation  of  the  liver  to  the 
metabolism  of  fat,  submits  the  following  lecithin  determinations  on 
the  livers  of  animals  under  known  conditions  of  feeding.  The 
amount  of  lecithin  in  the  liver  appeared  to  be  somewhat  nearly  con- 
stant, its  considerable  percentage  variations  being  due  in  large  part 
to  the  storage  or  removal  of  fat,  as  determined  by  the  general  state 
of  nutrition  of  the  animaL 


PHOSPHORUS  METABOLISM 


137 


CONTENT  OF  LECITHIN  IN  LIVER  OF  VARIOUS  ANIMALS  UNDER 
DIFFERENT  CONDITIONS  OF  FEEDING— Paton  (1895-96) 


Lecithin 

Animal 

Percent 
of  liver 

Percent  of 
solids  of  liver 

Percent  of  ether 
extract  of  liver 

Feed 

Rabbit 

2.60 
2.70 
2.05 
1.90 
2.57 
3.04 
3.31 
1.72 
1.42 
2.24 

11.4 
11.9 

9.1 
10.3 
11.7 
5.2 
9.8 
8.9 

56.1 

58.4 
51.3 
36.4 
50.0 
60.0 
70.0 
11.0 
3.53 
34.2 

Bran  and  oats 

Rabbit 

Rabbit 

Cat 

Kitten 

Kitten 

Fast  of  48  hours 

Kitten 

"     "  59      " 

Kitten 

Kitten 

Rabbit,  very  fat. . . 

Turnips 

Balthazard  (1901a)  investigated  the  lecithin  content  of  the 
liver  in  a  number  of  cases  of  different  diseases  or  artificially  pro- 
duced pathological  conditions  in  animals.  He  found  the  lecithin 
high  in  all  the  pathological  cases  examined;  thus  it  was  increased 
by  infection  in  tuberculosis  and  diphtheria,  by  intoxication  with  a 
mineral  poison,  phosphorus;  by  bacterial  poison — typhus  toxin,  or 
by  autointoxication,  as  in  inanition  and  uremia.  Balthazard  con- 
cluded that  a  large  part  of  the  hepatic  lecithin  comes  from  the  de- 
struction of  leucocytes  of  the  circulating  blood. 

The  livers  of  4  pigs  to  which  Hart,  McCollum  and  Fuller  (1909) 
had  given  different  amounts  and  different  kinds  of  phosphorus  in 
the  diet  showed  a  phosphorus  content  of  from  1.27  to  1.43  percent  P 
in  the  air-dry  substance,  with  an  average  of  1.35  percent. 

Forbes  (1909)  found  the  phosphorus  of  the  livers  of  pigs  to 
vary  between  0.291  and  0.367  percent,  and  the  phosphorus  in  the  ash 
between  24.18  and  28.23  percent,  as  affected  by  the  food  received. 

The  bile  and  liver  of  cattle  were  analyzed  by  Daniel-Brunet  and 
Rolland  (1911b)  and  the  phosphate  content  of  liver  showed  a  range 
from  2.90  to  3.48  parts  P205  per  1000  parts  fresh  substance  (1.26- 
1.52  parts  P). 

For  phosphorus  estimations  in  the  liver  in  various  pathological 
states  see  Robin  (1911). 

Partition  of  the  Phosphorus  in  Liver.  Plosz  (1873)  identified 
nuclein  in  liver,  and  A.  Kossel  (1882)  submitted  the  following  fig- 
ures for  nuclein  phosphorus  in  liver. 


TOTAL  PHOSPHORUS  AND  NUCLEIN  PHOSPHORUS  IN  LTVER 

Kossel  (1882)— Percent 

Species 

H3PO4    in    fresh    organ 

Nuclein  H3PO1 

Nuclein  H3PO4  in 
total  H3POi 

Dog    

0.846 
1.267 
1.077 
0.834 

0.444 
0.390 
0.511 
0.264 

52.5 

Cattle     

30.8 

Hen,  -well  fed.  .  .  . 

47.4 
31.7 

138 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


Drechsel  (1886)  showed  that  jecorin  is  found  in  liver.  Paul 
Manasse  (1895)  isolated  from  the  liver  of  the  horse  a  phosphatid 
which  yielded  grape  sugar  on  cleavage  and  hence  was  supposed  to 
be  jecorin.  Waldvogel  and  Tintemann  (1906)  also  report  jecorin 
in  the  liver.  Baskoff  (1908)  found  in  addition  to  lecithin  and 
jecorin  another  phosphatid  which  he  named  heparphosphatid.  Ken- 
naway  and  Leathes  (1909)  found  the  jecorin  fraction  of  the  lipoids 
from  the  liver  of  the  pig  higher  in  phosphorus  than  that  from  the 
liver  of  the  dog  or  the  goat. 

The  compounds  which  Liebermann  (1893a)  studied  and  called 
lecithalbumins  were  found  in  most  abundance  in  liver  and  lungs. 
Phosphocarnic  acid  is  reported  by  Siegfried  (1896)  and  by  Balke 
and  Ide  (1896).  Balke  and  Ide  give  the  amount  as  0.183  percent 
in  the  liver  of  the  dog  and  0.015  percent  in  that  of  the  horse. 
Levene  and  Mandel  (1908b)  say  that  among  the  nucleic  acids  is 
guanylic  acid,  and  others  describe  a  ferruginous  nucleoprotein,  or 
mixture  of  such  nucleoproteins,  to  which  the  name  "ferratin"  is 
given. 

Masing  (1911b)  published  a  series  of  analyses  which  indicate  a 
percentage  decrease  in  amount  of  nucleic  acid  in  the  rabbit  and  in 
rabbit  livers  with  advancing  stages  of  development,  both  before  and 
after  birth. 

NUCLEIN-PHOSPHORUS  IN  RABBIT  EMBRYOS  AND  RABBIT  LIVERS 

Masing  (1911) 


Stage  of  development 


18  embryos  0.5—1.5  cm.  long  from  1st  half  of 

gestation;  total  N  about  21  mg 

2  embryos  averaging  21.5  gm.  in  weight;  about  the 
beginning  of  the  4th  week  of  gestation 

1  embryo  at  same  stage  as  the  last;  considerably 

less   N 

2  embryos  somewhat  older 

5  livers  from  stage  somewhat  later  than  the 

other    livers    

4  embryos   (5  livers)  about  1-2  days  before  birth. 

2  embryos,  mature 

Young  just  after  birth  (small  breed;  4  livers) 
11  days  old  animal  (skin  and  stomach  contents 

removed)    

2  livers  of  11  days  old  animals   

2  livers  of  22  days  old  animals 

Liver  of  a  full-grown  animal 

Ditto     


Average 

Nuclein  P 

Average 

weight  of 

of  whole 

weight  of 

animal 

animal 
for  0.35 
gm.  N 

liver 

Grams 

Milli- 
grams 

Grams 

20.3 

21.5 

17.8 

2 

22.5 

17. 

28. 

14.7 

28 

1.8 

36 

13. 

2.6 

43 

12. 

33 

11.7 

2.3 

57 

11.9 

72 

4.0 

210 

9.5 

1800 

65. 

Nuclein  P 
of  liver 
for  0.35 

gm.  N. 

Milli- 
grams 


22.8 


20.4 
18.0 


17. 


16. 
12 
11.5 
10. 


PHOSPHORUS  METABOLISM 


139 


Scaffidi  (1908)  made  nucleoprotein  determinations  on  the  livers 
of  rabbits  in  'connection  with  his  study  of  the  iron  distribution  in 
the  liver  after  feeding  an  iron  paranucleinate  called  "triferrin,"  This 
paranucleinate  did  not  affect  either  the  nucleoprotein  content  of  the 
liver  or  the  phosphorus  content  of  the  nucleoprotein,  as  it  did  ap- 
parently the  iron.  The  amounts  of  nucleoprotein  found  in  the  livers 
of  the  several  rabbits  range  from  0.3595  to  1.2334  percent  of  the 
liver,  and  the  phosphorus  content  of  the  nucleoprotein  from  2.51  to 
2.83  percent,  while  the  iron  content  of  the  nucleoprotein  in  the  con- 
trol animals  ranged  from  0.18  to  0.44  percent,  averaging  0.30  per- 
cent, and  that  of  the  experiment  animals  from  0.34  to  1.10  percent, 
averaging  0.67  percent. 

PHOSPHORUS  IN  SPLEEN 
Phosphorus  Content.  Kriiger  (1895)  examined  spleens  and 
livers  of  cattle  fetuses,  calves  and  adults .  The  phosphorus  content 
of  the  spleen  was  found  to  be  highest  when  the  fetus  was  30-60  cm. 
in  length,  and  lowest  in  the  adult  animal.  In  the  adults,  livers  and 
spleens  gave  about  the  same  figure. 

AVERAGE  PHOSPHORUS  (P)  CONTENT  OF  THE  SPLEEN  OF  CATTLE 
OF  DIFFERENT  AGES  (Kriiger,  1895)— Percent  Dry  Basis 


Fetuses,  length  in  centimeters 

Calves 

Cows 

Oxen 

30-40 
2.38 

40-50 
2.43 

50-60 
2.39 

60-70 
2.13 

70-80 
1.94 

80-90 
1.70 

90-100 
1.48 

1  .82 

1.26 

1.37 

Dhere  and  Maurice  (1910)  find  a  steady  decrease  of  phospho- 
rus in  the  spleen  of  dogs  with  advancing  age. 

INFLUENCE  OF  AGE  ON  THE  QUANTITY  AND  PARTITION  OF  PHOS- 
PHORUS IN  THE  SPLEEN  OF  DOGS 
Dhere  and  Maurice  (1910) 


Num- 
ber of 
dogs 

Phosphorus, 
percent  of 

fresh 
substance 

Phosphorus, 
percent  of 

dry- 
substance 

Percent  of  total  phosphorus 

Age 

Lipoid 

Nucleic 

Inorganic 

Few  hours  to  15  days 

21 
9 
6 
4 

0.35 
0.34 
0.29 
0.25 

1.70 
1.61 
1.32 
1.09 

13.55 
16  75 

8.67 

7  85 

77  78 

4  weeks  to  4  months 

75  40 

6  months  to  15  months 

16.91            6.57 
21  17          ii  ns 

76.52 
67  78 

Partition  of  the  Phosphorus  of  Spleen.  A.  Kossel  (1882)  sub- 
mitted the  following  figures  for  total  and  nuclein  phosphorus  in 
spleen. 


140  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

TOTAL  AND  NUCLEIN  PHOSPHORUS  IN  SPLEEN— (Kossel,  1882)  Percent 


Species 

H3P04  in  fresh 
organ 

Nuclein  H3PO4 

Nuclein  H3PO4  in 
total  H3PO4 

Cattle    

1.162 

0.837 
1.005 

0.867 
0.593 
0.636 
0.676 

74.6 
70.9 
63.3 
67.3 

J- Duplicates 

Liebermann  (1893a)  found  considerable  of  lecithalbumin  in 
spleen.  Gourlay  (1894)  isolated  from  spleen  a  substance  he  called 
nucleoalbumin.  Phosphocarnic  acid,  phosphatids  and  nucleoproteins 
are  found.  Besides  the  jecorin  (Waldvogel  and  Tintemann,  1906), 
which  is  free  from  iron,  Burow  (1910)  found  three  iron-containing 
phosphatids  in  the  spleens  of  cattle  and  of  men.  The  presence  of 
iron  in  the  lipoids  of  spleen  is  considered  significant  in  connection 
with  the  view  that  the  chief  physiological  function  of  the  spleen, 
aside  from  leucocyte  formation,  may  be  iron  metabolism.  Capez- 
zuoli  (1909a)  found  considerable  iron  in  the  nucleoprotein.  Ac- 
cording to  Sato  (1909),  the  iron  of  the  nucleoprotein  is  present  in 
two  forms,  one  more  stable  than  the  other. 

Sato  (1909)  obtained  0.709,  0.406,  0.481,  and  0.498  percent  nu- 
cleoprotein from  the  spleens  of  cattle.  Corper  (1912)  reports  that 
the  spleen  of  the  dog  contains  6-7  percent  lecithin;  about  0.27-0.52 
percent  water-soluble  phosphorus  and  about  0.39  percent  insoluble 
phosphorus.  _1_ 

Panella  (1904c)  made  water  and  nucleon  determinations  on  the 
fresh  spleen  of  cattle,  horses,  sheep,  swine  and  dogs,  and  found  it. 
present  in  all  in  amounts  varying  somewhat  with  the  species.  The 
nucleon  content  of  the  spleen  of  cattle,  sheep  and  dogs  seems  to  de- 
crease with  increasing  years. 

The  values  found  are  as  follows : 

WATER  AND  NUCLEON  CONTENT  OF  FRESH  SPLEEN— Percent 


Animal 

Water 

Nucleon 

Fresh  subs. 

Dry  subs. 

Cattle     ' 

78.88 
78.64 
77.27 
78.92 
78.79 

0.31 
0.36 
0.40 
0.57 
0.67 

1.46 

1.17 

1.76 

2.70 

3.16 

PHOSPHORUS  IN  THE  PANCREAS 
Phosphorus  Content.     Liining  (1899)  reports  the  phosphoric 
acid  of  the  ash  of  the  pancreas  of  two  aged  women  as  46.99  and 
45.29  percent,  respectively.     See  also  Gossmann  (1898)  and  Juchler 
(1912). 


PHOSPHORUS  METABOLISM 


141 


Juchler  (4913)  found  phosphorus  in  the  pancreas  of  adults  in 
amounts  varying  within  narrow  limits,  apparently  mainly  in  organic 
condition.  The  percentage  of  total  P205  in  the  fresh  pancreas  at 
various  ages  is  as  follows :  New-born  2.36 ;  7  mos.-5  years  1.03 ;  20 
years-48  years  0.55 ;  50  or  more  years  0.42 ;  adults  in  general  0.48. 

Partition  of  the  Phosphorus.  Frankel,  Linnert  and  Pari 
(Frankel  and  Pari,  1909;  Frankel,  Linnert  and  Pari,  1909)  have 
made  a  special  study  of  a  phosphatid  found  in  the  pancreas  of  cattle. 

Plimmer  and  Kaja  (1909)  show  that  a  prominent  feature  in  the 
phosphorus  changes  in  the  pancreas,  accompanying  the  secretion  of 
pancreatic  juice,  is  the  destruction  of  phosphoprotein,  which  de- 
creases from  about  2.5  percent  of  the  total  phosphorus  to  less  than 
half  of  one  percent  of  the  total  phosphorus.  The  total  phosphorus 
of  the  pancreatic  juice  is  very  small  in  amount.  Its  total  protein 
phosphorus  is  phosphoprotein,  and  inorganic  phosphorus  is  also 
present.  It  contains  no  lecithin.  Below  are  the  analytical  data  on 
this  matter. 

PHOSPHORUS  (P205)  CONSTITUENTS  OF  PANCREAS  AND  PANCRE- 
ATIC JUICE— Percent 


Condition  of 
Phosphorus 

Dog's  pancreas,  normal 

Dog's  pancreas 

after  action  of 

secretin 

Pancreatic  juice 
of  dog 

I 

II 

III 

IV 

I 

II 

III 

I 

II 

III 

Ether  soluble 
(lecithin) 

Water  soluble 
(nucleic  acid-(- 
inorganic)     .  . 

Protein   (nucleo- 
protein -(-phos- 
phoprotein) 

Phosphoprotein   . 

8.6 

37.6 
16.4 

53.8 
2.5 

28.5 

15.5 
7.1 

56.0 
2.6 

30.6 

17.4 
7.8 

52.0 
1.6 

28.6 

14.4 

6.4 

57.0 
1.9 

31.1 

16.2 
7.3 

52.7 
0.4 

28.0 

19.8 
6.9 

52.2 
0.3 

31.0 

16.2 

6.7 

52.8 
0.0 

0.0 

56.3 

present 

43.7 
43.7 

0.0 

86.3 
trace 

13.7 
13.7 

0.0 

59.8 
trace 

36.7 
40.2 

A.  Kossel  (1882)  reports  the  total  phosphorus  (H3P04)  of  the 
pancreas  of  cattle  as  1.257  and  1.215  percent,  the  nuclein  phospho- 
rus 0.580  and  0.606  percent,  and  the  percent  of  nuclein  phosphorus 
in  the  total  phosphorus  as  46.1  and  49.9  percent.  These  pairs  of 
figures  are  duplicate  determinations. 

PHOSPHORUS  IN  KIDNEY 

Phosphorus  Content.  From  Gossmann  (1898)  we  compute  the 
percentages  of  phosphoric  acid  in  the  kidneys  of  man  and  steer,  as 
follows :  man  0.107  percent ;  steer  0.401  percent. 

Forbes  (1909)  found  in  the  kidneys  of  swine  phosphorus  vary- 
ing between  0.205  and  0.318  percent  and  in  the  ash  18.47-27.90  per- 
cent. 


142 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BTJL.  5 


Partition  of  the  Phosphorus.  A.  Kossel  (1882)  found  in  bovine 
kidney  0.7584  percent  H3P04,  0.285  percent  nuclein  H3P04  and  37.6 
percent  nuclein  P  in  the  total  P. 

Lonnberg  (1890)  reports  finding  nucleoalbumins  in  the  cortical 
substance  and  the  medullary  substance  of  kidney,  and  in  the  mucous 
membrane  of  the  urinary  bladder.  Liebermann  (1891b,  1893a, 
1893b)  found  in  the  parenchyma  of  the  kidney  a  compound  which  he 
called  lecithalbumin,  and  which,  after  removal,  had  such  properties 
as  to  suggest  that  in  the  tissues  it  might  be  the  agent  by  which  the 
kidney  excretes  acid  urine  from  alkaline  blood.  The  substance  in 
the  kidneys  which  others  had  called  protagon  Panzer  (1906)  states 
to  be  a  cholesterol  compound  not  containing  phosphorus.  Balke  and 
Ide  (1896)  obtained  0.043  percent  phosphocarnic  acid  from  the  kid- 
ney of  the  horse  and  0.205  percent  from  that  of  the  dog.  Lusena 
(1903)  gives  the  lecithin  content  of  the  kidneys  of  rabbits  as  aver- 
aging 2.086  percent  for  the  normal  organs  and  1.856  for  organs  af- 
fected by  experimental  fatty  degeneration.  Rubow  (1905)  reported 
the  following  lecithin  phosphorus  figures  for  the  kidneys  of  dogs  in 
various  conditions. 

LECITHIN  AND  FAT  CONTENT  OF  THE  KIDNEYS  OF  DOGS 
Rubow  (1905)  Percent,  Fresh  Basis 


Condition  of  dog 

Kidney 

Ether  extract 

Lecithin 

Fat 

j               Right 
1                Left 
j               Right 
1                Left 
j  Right  extirpated 
1               Left 
J    Left  extirpated 
1              Right 

j    Left  extirpated 
1              Right 
j    Left  extirpated 
1              Right 

13.81 
13.63 
15.21 
14.99 
19.46 
21.28 
14.06 
14.97 
15.04 
15.74 
13.61 
13.13 
15.65 
15.79 

12.49 

15.33 

6.60 
6.86 
8.66 
8.63 
7.57 
7.95 
7.87 
7.39 
8.55 
8.32 

7.89 

7.90 

12.86 

14.42 
5.40 
6.34 
7.47 

Phosphorus-poisoned,  about  3  mg. 

per  kg.  body  weight,  injected.. 
Phosphorus-poisoned,  about  3  mg. 

per  kg.  body  weight,  injected.. 
Under   chloroform  for  5  hrs.  two 

7.79 
5.74 
5.74 
7.10 

7.47 

4.60 

Under  chloroform  for  5  hrs.  two 

7.43 

Dunham  (1903-4,  1904-05,  1905-06,  1908)  and  Dunham  and 
Jacobson  (1910)  isolated  and  studied  a  peculiar  phosphatid  found  in 
the  kidneys  of  cattle,  which  they  call  "carnaubon."  Frankel  and 
Nogueira  (1909a,  1909b)  describe  three  unsaturated  phosphatids. 
MacLean  (1912a,  1912b,  1912-13)  has  made  painstaking  examina- 
tions of  the  phosphatids  of  the  kidney  of  the  horse  and  finds  the 
chief  one  to  be  a  lecithin,  or  at  least  a  phosphatid  having  the  N:P 
relation  1:1.  He  finds  also  cuorin  and  a  diamino-monophosphatid 
which  he  considers  as  probably  the  carnaubon  of  Dunham  and  Ja- 
cobson freed  from  a  nitrogenous  impurity  present  in  their  product. 


PHOSPHORUS  METABOLISM 


143 


PHOSPHORUS  IN  SUPRARENAL  CAPSULES 

Paul  Mariasse  (1895)  isolated  from  the  adrenals  of  the  horse 
,and  of  cattle  a  sugar-yielding  phosphatid  supposed  to  be  jecorin. 
Orgler  (1904)  reports  a  substance  similar  to  the  so-called  protagon 
of  the  brain  (0.6  percent  of  the  substance) ,  and  Rosenheim  and  Tebb 
(1909b)  found  in  the  adrenals  a  small  amount  of  the  sphingomyelin 
such  as  they  consider  a  prominent  constituent  of  the  protagon  of 
brain. 

Bernard,  Bigart  and  Labbe  (1903a,  1903b)  identified  as  lecithin, 
or  a  mixture  of  lecithins,  the  distinctive  fat  occurring  in  the  spong- 
iocytes  of  the  suprarenal  capsules  and  increasing  during  the  func- 
tioning of  the  organs.  It  is  looked  upon  as  a  secretion  of  these 
cells.  The  phosphorized  fat  was  found  to  be  in  the  horse  45.3  per- 
cent, in  the  sheep  48.8  percent,  in  the  rabbit  52.7  percent  and  in  man 
(one  case)  13.1  percent  of  the  total  fat  of  the  organ.  The  total 
amount  found  was  in  the  horse  6.77  percent  of  the  gland  and  in  man 
2.08  percent. 

The  following  are  Alexander's  (1892)  determinations  of  the 
lecithin  and  the  nuclein  phosphoric  acid. 

LECITHIN  AND  NUCLEIN  PHOSPHORIC  ACID  DETERMINATIONS  ON 
THE  ADRENALS  OF  THE  HORSE  (Carl  Alexander,  1892) 


Age  of 

Lecithin 

Nuclein  phosphoric  acid 

horse 

Percent   of  fresh 
substance 

Percent   of   dry 

substance 

H3POt,   percent  of 
fresh  substance 

H3PO4,  percent 
of  dry  substance 

16   years.  . 
3    years . . 
2    years . . 
2    years . . 

4.2973 
2.81383 
2.573 
3.4502 

26.018 
16.6114 
16.071 
21.964 

0.7781 

4.592 

PHOSPHORUS  IN  THE  THYMUS 

Lilienfeld  (1894)  reports  a  general  chemical  examination  of 
leucocytes  from  the  thymus  of  calves.  He  found  a  nucleoprotein, 
protagon,  inosite,  monopotassium  ^  phosphate,  lecithin  and  nucleo- 
histone.  The  total  phosphorus  is' given  as  3.01  percent  of  the  dry 
substance,  the  lecithin  as  7.51  percent  and  the  nuclein  as  68.78  per- 
cent. 

Huiskamp  (1901a)  says  that  69.4  percent  of  the  proteins  of  the 
thymus  are  in  nucleohistone  and  18.7  percent  in  nucleoprotein,  leav- 
ing only  11.9  percent  of  other  proteins.  Malengreau  (1900,  1901) 
states  that  there  are  two  nucleoalbumins  and  two  histones  in  the 
thymus.  Herlitzka  and  Borrino  (1902,  1902b,  1903)  try  to  distin- 
guish the  parts  played  by  the  nucleohi  stones  and  the  nucleoproteins 
of  the  thymus,  the  liver  and  the  kidney. 


144 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


The  nucleohistone  of  the  thymus  has  been  given  considerable 
attention.  (See  Lilienf  eld,  1894, 1895 ;  Bang,  1900a,  1900b ;  Kossel, 
A.,  1900 ;  Malengreau,  1901 ;  Huiskamp,  1901a,  1901b,  1903;  Herlitz- 
ka  and  Borrino,  1902a,  1902b,  1903).  A  nucleohistone  is  a  nucleo- 
protein  in  which  the  nucleic  acid  is  united  with  one  of  the  simple 
proteins,  a  histone.  The  result  is  a  nucleoprotein  differing  from 
most  of  the  members  of  this  group  through  the  content  of  a  larger 
proportion  of  nucleic  acid  and  relatively  little  protein.  According 
to  Goubau  (1911)  it  is  characteristic  of  little-differentiated  cells. 
There  seems  to  be  present  in  the  thymus  at  least  one  nucleoprotein 
of  this  type  and  one  of  the  type  containing  relatively  less  nucleic 
acid.  It  will  be  observed  that  a  very  large  proportion  of  the  proteins 
of  this  gland  are  united  with  phosphorus  in  one  or  the  other  of  these 
ways.  Goubau  finds  nucleohistone  much  more  abundant  in  thymus 
than  in  any  other  mammalian  organs,  and  says  that  this  nucleohis- 
tone of  the  thymus  is  of  a  different  type  from  that  of  the  other  or- 
gans ;  it  is  insoluble  in  physiological  salt  solution.  It  is  not  found  in 
human  or  horse  serum  nor  in  ascitic  fluid  containing  bile.  It  is  prob- 
able that  the  histones  and  nucleohistones  are  fixed  in  such  organs 
as  the  thymus  and  the  spleen,  and  hence  cannot  be  carried  away  in 
body  fluids. 

PHOSPHORUS  IN  THE  THYROID 

According  to  Gourlay  (1894),  the  only  protein  of  the  thyroid 
is  a  nucleoalbumin  which,  from  microchemical  examination,  he  con- 
cluded is  derived,  at  least  in  part,  from  the  colloid  matter  of  the 
acini. 

Baldoni  (1899-1900)  gives  these  phosphorus  and  nucleoprotein 
determinations  on  thyroids  of  different  animals. 


ANALYSES  OF  THYROID  GLANDS— (Baldoni,  1900)  Percent 

Species 

Water 

Ash 

P2O5  in  ash 

P2O5  in  fresh 
substance* 

Nucleoprotein 

in  dry 

substance* 

Nucleoprotein 

in  fresh 

substance 

68.09 
74.71 
69.79 
70.81 
73.30 
73.79 

0.92 
0.74 
1.00 
1.03 
0.92 
0.98 

12.59 
23.24 
12.48 
13.59 
14.66 
16.31 

0.116 
0.172 
0.125 
0.140 
0.135 
0.160 

5.55 
3.49 
4.15 
3.43 
3.69 
4.45 

1.771 

Cattle  (Tuscan).. 
Cattle  (Maremma) 
Buffalo 

0.883 
1.254 
1.001 

0.985 

1.166 

*   Computed  by  F.  &  K. 

Markotum  (1895)  describes  from  the  thyroid  of  cattle  a  phos- 
phorus-containing substance  "thyreonucleoalbumin,"  which  is  said 
to  have  the  following  composition:  C  51.46,  N  15.56,  H  6.94,  P  0.32, 
S  1.5  and  O  24.12.  Markotum  says  that  the  alteration  of  the  nerv- 
ous system  by  the  thyroid  is  supposed  to  be  through  this  compound. 


PHOSPHORUS  METABOLISM 


145 


Oswald  (1899)  finds  that  the  active  agent  of  the  thyroid  gland 
as  used  in  pathological  cases,  the  so-called  thyroid  gland  colloid,  is 
a  mixture  of  two  compounds.  The  iodine-containing  body,  thyreo- 
globulin, which  was  shown  to  have  the  characteristic  influence  on 
nitrogen  metabolism  when  the  gland  was  administered  to  dogs,  is 
free  from  phosphorus;  but  it  is  associated  in  the  colloid  with  a 
small  amount  of  nucleoprotein  containing  0.16  percent  phosphorus. 

Fenger  (1913),  in  connection  with  an  extensive  study  of  the 
iodine  content  of  the  thyroid  glands  of  animals  in  both  the  adult  and 
the  fetal  stage,  has  found  with  regard  to  cattle  thaf'enlarged  glands 
in  general,  both  fetal  and  adult,  contain  less  total  iodine  and  much 
more  total  phosphorus  than  normal  thyroids/'  and  that  "normal 
fetal  glands  are  relatively  larger  and  contain  more  iodine  and  phos- 
phorus per  unit  of  body  weight  than  thyroids  from  fully  mature  an- 
imals." 

In  a  later  paper  Fenger  (1914)  reported  studies  on  the  com- 
position of  the  thyroid  gland  in  cattle  as  affected  by  sex  and  preg- 
nancy.     Among  his  conclusions  are  the  following : 

"Castrated  males  contain  less  thyroid  tissue  than  either  un- 
castrated  males  or  females,  but  the  iodine  content  per  unit  of  body 
weight  is  about  half-way  between  the  uncastrated  male  and  female 
animals. 

"The  phosphorus  content  of  the  thyroid  gland  seems  to  be  fair- 
ly uniform  in  all  four  cases  and  should  only  be  considered  as  indi- 
cation of  normal  physiological  activity." 

From  the  analytical  data  we  select  the  following: 

PHOSPHORUS    AND    IODINE    CONTENT    OF    THYROID    GLAND    OP 

CATTLE 


Phosphorus  in 
fresh  gland 

Mg. 

Iodine  in  fresh 
gland 

Mg. 

Fresh  gland 

tissue  per  100 

lbs.  body 

weight 

Grams 

Phosphorus 
per  100  lbs. 
body  weight 

Mg. 

Iodine  per 

100  lbs.  body 

weight 

Mg. 

Bulls    

Pregnant  cows 
Non-pregnant 
cows 
and  heifers . . 

30.66 
23.97 
21.08 

20.52 

19.38 
19.73 
18.50 

16.98 

2.14 
1.92 

2.44 

2.44 

2.19 
2.00 
2.11 

2.22 

1.38 
1.64 
1.85 

1.84 

PHOSPHORUS  IN  THE  LUNGS 

By  Sieber  and  Dzierzgowski  (1909)  the  mixed  lung  tissue  of 
several  horses  was  found  to  contain  76.6  percent  water,  and  of  the 
dry  substance  1.107  percent  was  phosphorus;  which  equals  0.259 
percent  in  the  fresh  substance. 


146  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

The  compounds  which  Liebermann  (1893a)  studied  and  called 
lecithalbumins  were  found  abundant  in  lungs.  According  to  Sieber 
(1909b),  the  phosphatid  extract  of  lungs  (horse)  includes  lecithin 
and  jecorin  in  the  approximate  proportion  6:1.  A  cholesterol-like 
substance  was  also  isolated.  There  is  a  discussion  by  Zoja  (1894) 
on  the  lecithin  of  the  alveolar  cells  of  the  lungs  and  the  semeiological 
significance  of  the  myelin  drops  of  the  sputum. 

PHOSPHORUS  IN  THE  HYPOPHYSIS 

According  to  the  analyses  of  Malcolm  (1904),  the  pituitary 
gland  contains  in  its  "glandular  portion"  0.72  percent  P  and  in  its 
"nervous  portion"  0.8  percent. 

PHOSPHORUS  IN  THE  DIGESTIVE  MUCOSA 

After  peptic  digestion  of  the  mucous  membrane  of  the  stomach, 
Liebermann  (1891a)  obtained  a  lecithin-albumin  complex  which  he 
thought  existed  as  such  in  the  walls  of  the  stomach  and  there  func- 
tioned significantly  in  the  control  of  stomach  acidity. 

Borri  (1906)  studied  the  phosphorus  compounds  of  the  mucous 
membrane  of  the  intestine  (dog) ,  distinguishing  nucleoprotein, 
phosphorized  fats,  lecithalbumin  and  perhaps  some  other  phos- 
phorized  organic  body  as  well  as  inorganic  phosphate.  He  says  that 
there  are  no  nucleoalbumins  present.  The  phosphate  content  of  a 
water  extract  of  the  membrane  was  distinctly  less  after  fast.  The 
ingestion  of  oleic  acid  after  a  three-days'  fast  increased  the  phos- 
phate content  of  the  extract,  which  is  interpreted  as  supporting  the 
idea  that  glycerin  to  unite  with  the  fatty  acid  was  obtained  from 
the  lecithalbumin  of  the  membrane. 

Araki  (1903b)  isolated  nucleic  acid  from  the  mucous  membrane 
of  the  small  intestine  to  the  extent  of  0.66  percent. 

PHOSPHORUS  IN  THE  GENITALIA 

Paton  and  his  associates  (Paton,  1898;  Paton,  et  al., 
1897-98),  in  their  extensive  study  of  the  metabolism  of 
salmon  in  fresh  water,  gave  considerable  attention  to  the 
forms  and  amounts  of  phosphorus  present  in  the  genitalia 
and  muscles  of  salmon  at  the  time  of  leaving  the  sea  in 
May,  and  again  in  October  after  the  long  fast  in  fresh  water,  during 
which  time  the  reproductive  organs  and  products  mature.  The 
muscles  were  shown  to  have  the  phosphorus  most  largely  present 
as  inorganic  phosphates,  while  the  ovaries  had  greater  amounts  of 
lecithin  and  of  phosphoprotein  (ichthulin) ,  and  the  testes  had  more 
than  a  half  of  it  as  nuclein.  The  muscles  lose  phosphorus  while  the 
genitalia  gain  it.  The  following  are  given  as  representative  figures 
showing  the  distribution  of  phosphorus  in  each  of  the  three  classes 
of  tissue. 


PHOSPHORUS  METABOLISM 


147 


DISTRIBUTION  OF  PHOSPHORUS  IN  THE  MUSCLES  AND  GENITALIA 

OF  SALMON  (Paton,  1897-98)— Percent 


Tissue 

No.  of 
fish 

In  ether 
extract 
(lecithin) 

In  water 

extract 

(inorganic 

phosphates 

In  residue 

(nucleins  or 

pseudo- 

nucleins) 

Total 

Muscle,  thick . . 
Muscle,  thin. . . 

Muscle,  thick.. 

Muscle,  thin. . . 

14 
14 

76 

76 
14 

76 
53 
68 

0.0416 
0.0460 

0.060 

0.060 
0.114 
0.150 
0.063 
0.060 

0.131 
0.094 

0.095 

0.119 

.      0.057 
0.075 
0.068 
0.040 

0.056 
0.041 

0.055 

0.063 
0.114 

0.189 
0.161 
0.178 

0.228 
0.181 

0.210 

0.242 

1.285 
0.414 
0.292 
0.278 

Estuary  fish 

The  "thin"  muscle  is  said  to 
make  up  about  K  of  the  body 
Estuary  fish  taken    later  in 
the  year 

Testis 

Testis 

Milroy  (1908)  has  since  made  a  similar  study  of  herring.      The 
percentage  content  of  phosphorus  (P205)  in  ovaries,  and  the  weights 
of  phosphorus  per  average  fish  found  in  these  organs  are  averaged 
for  the  months  of  the  year  as  follows : 
PHOSPHORUS  (P2O5)  IN  GENITALIA  OF  THE  HERRING— (Milroy,  1908) 


3 

£ 

a 

fao 

Condition 

3 

3  a 

OOP 

til 

OQ 

OP 

.  0 
>  a 

PIS 

0  ctf 
£  ft 
Pen 

is 

+> 

£>  a; 

focc 

Ovaries 

P2O5,  percent 

077 

1.23 

0.71 

0.93 

0.91 

1.03 

1.19 

0.93 

0.91 

P2O5,  per  fish 

0.003 

0.011 

0.01 

0.05 

0.26 

0.48 

0.34 

0.16 

0.016 

Testes 

P2O5,  percent 
P2O5,  per  fish 

0.86 
0.20 

2.10 
1.21 

Plimmer  and  his  associates  (Plimmer  and  Scott,  1908 ;  Plimmer 
and  Kaya,  1909),  in  their  study  of  the  distribution  of  phosphopro- 
teins  in  animal  products,  found  them  chiefly  in  milk,  egg-yolk  and 
the  ova  of  fish ;  but  they  were  present  also,  in  smaller  proportions, 
in  the  pancreas  and  salivary  glands.  They  do  not  appear  in  the 
thymus  nor  in  the  testes  of  the  bull  or  the  codfish.  Determinations 
were  made  also  of  the  proportionate  amounts  of  the  different  kinds 
of  phosphorus  compounds  in  the  testes  of  codfish  and  in  the  eggs  of 
the  frog. 

DISTRIBUTION  OF  PHOSPHORUS  IN  CERTAIN  TISSUES 
Plimmer  and  Kaya  (1909)— Percent  of  Total  P205 


Water  sol- 

Protein 

Ether 

uble  (nucle- 

(nucleo-pro- 

Substance 

Animal 

soluble 

ic  acid  and 

Inorganic 

tein  and 

Phospho- 

examined 

(lecithin) 

inorganic) 

phospho- 
protein) 

protein 

Codfish 

17.8 

22.6 

20.5 

59.6 

,   0     . 

Codfish 

8.9 

31.9 

20.4 

59.2 

0 

Eggs,   ovarian .... 

Frog 

26.2 

4.3 

0 

69.5 

61.9 

Eggs,   soon   after 

being  laid   .... 

Frog 

19.1 

13.0 

Trace 

67.9 

41.1 

Eggs,  soon  after 

being  laid   .... 

Frog 

21.4 

9.6 

Trace 

69.0 

40.9 

148  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Panella  (1903d,  1903i,  1904a)  found,  of  phosphocarnic  acid, 
1.1733  percent  in  the  dry  testicular  substance  of  the  ass,  and  0.871 
percent  in  that  of  the  horse. 

Konig  and  Grossfeld  (1913a),  in  considering  the  food  value  of 
fish  sperm  mention  the  high  phosphorus  content  (4.46  percent  of 
the  ash-free,  dry  substance  of  herring  sperm  and  3.74  percent  of 
that  of  carp  sperm),  and  the  high  content  of  lecithin  (20.7  per- 
cent of  the  fat  of  herring  sperm  and  20.2  percent  of  that  of  carp 
sperm).  In  a  second  article  (1913b)  they  report  the  phosphorus 
content  of  the  ichthulin  and  albumin  and  the  lecithin  content  of  the 
fat  of  the  roe  of  several  species  of  fish.  The  dry,  ash-free  ichthulin 
from  herring  roe  contains  1.56  percent  and  the  albumin  1.05  percent 
phosphorus,  while  the  lecithin  content  of  the  fat  of  herring  roe  was 
reported  as  43.61  percent. 

Rosenbloom  (1913a)  reports  determinations  of  the  percent- 
ages of  lipins,  phospholipins,  neutral  fat,  fatty  acids  and  cholesterol 
in  the  ovary  and  corpus  luteum  of  the  cow  in  the  non-pregnant  state, 
and  in  the  pregnant  state.  The  results  indicate  that  there  is  prac- 
tically no  increase  in  the  above  substances  during  pregnancy  in  the 
cow.  The  phospholipins  amount  to  3.94  percent  of  the  dry  sub- 
stance in  the  non-pregnant  ovary  and  3.48  and  3.63  percent  at  two 
stages  of  pregnancy ;  those  of  the  non-pregnant  corpus  luteum  were 
14.10  percent,  and  of  the  pregnant,  14.90  and  14.87  percent  of  the 
dry  substance. 

Bienenfeld  (1912),  Ballerini  (1912)  and  Sakaki  (1912, 1913a,  b, 
c  and  d)  have  reported  special  lipoid  studies  on  the  placenta.  Sakaki 
isolated  two  phosphatids,  sphingomyelin  and  jecorin. 

Spermatozoa  heads,  being  mainly  nucleus,  are  composed  largely 
of  nucleic  acid,  and  this  is,  in  the  case  of  several  fish  at  least,  com- 
bined with  protamin  or  histone  in  a  compound  more  simple  than 
most  nucleoproteins  of  the  animal  body  or  its  products.  This  relation 
has  been  especially  studied  by  Miescher  (1878,  1896,  1897)  and  by 
Mathews  (1897).  The  heads  are  apparently  free  from  lipoids,  but 
the  tails  contain  considerable  amounts,  and  largely  as  a  phosphatid 
which  is  reported  as  lecithin.  Gobley  (1850b)  also  found  the  leci- 
thin here.  Mathews  reports  that  jecorin  is  not  present.  The  ash 
of  spermatozoa  is  said  to  be  mainly  (three  quarters)  potassium 
phosphate.  The  seminal  fluid  contains  nucleon,  and  its  mineral 
matter  is  chiefly  calcium  phosphate  and  sodium  chloride. 


PHOSPHORUS  METABOLISM  149 

Percival-(1902)  made  phosphorus  studies  on  a  number  of  ani- 
mal organs.  The  total  phosphorus  figures  on  the  oxidized  tissue 
are  as  follows,  being  given  as  percent  of  P203  in  the  fresh  substance, 
and  arranged  in  order  of  decreasing  amount  of  phosphorus :  Thymus 
1.223;  heart  (sheep)  1.011;  pancreas  (sheep)  0.749;  lung  (sheep) 
0.745 ;  brain  (sheep)  0.635 ;  spleen  (beef)  0.570 ;  liver  (beef)  0.561 ; 
testicles  of  calf  0.517;  muscles  (beef)  0.506;  testicles  of  bull  0.470; 
kidney  (sheep)  0.454;  ovary  (cow)  0.429;  udder  (cow)  0.414;  thy- 
roids (sheep)  0.369. 

For  the  phosphorus  content  of  the  ether  extract  of  many  gland- 
ular organs  see  Gerard  and  Verhaeghe  (1911). 

PHOSPHORUS  OF  THE  BLOOD 

Kinds  of  Phosphorus  Compounds  Present.  The  blood  was, 
naturally,  early  recognized  as  an  important  part  of  the  animal  or- 
ganism the  chemical  composition  of  which  might  be  of  great  signi- 
ficance. In  some  cases  the  blood  was  ashed,  and  determinations  of 
mineral  constituents  made  on  the  ash ;  but  in  other  cases  the  indi- 
vidual organic  constituents  were  noted.  Ash  analyses  are  not  very 
enlightening,  and  quantitative  determinations  of  the  individual  com- 
pounds cannot  be  interpreted  with  definiteness  unless  the  corpuscles 
and  serum  be  examined  separately. 

Phosphorus  is  present  as  inorganic  salts,  chiefly  potassium 
phosphate  and  calcium  phosphate,  and  in  organic  combination  as 
phosphatids  and  nucleoproteins  or  nucleins,  and,  according  to  Pan- 
ella,  as  phosphocarnic  acid. 

In  1851  Gobley  (1851)  found  lecithin  in  blood.  "Protagon" 
studies  were  reported  by  both  Herrmann  (1866)  and  Hoppe-Seyler 
(1866a).  One  of  the  first  findings  of  nucleoprotein  was  in  blood 
corpuscles  (Plosz,  1871).  Pribram  (1871)  made  phosphorus  and 
calcium  determinations  which  distinguished  between  the  amounts 
precipitable  directly  from  the  defibrinated  and  centrifuged  blood 
serum  and  that  obtained  in  precipitable  form  after  ashing,  or,  as  it 
may  be  interpreted,  between  the  calcium  and  phosphorus  present  in 
inorganic  form  and  the  total  present.  The  indication  was  that  none 
of  the  calcium  was  in  organic  union,  while  more  than  two-thirds  of 
the  phosphorus  was  in  such  a  form.  Fokker  (1873)  reported  find- 
ing calcium  acid  phosphate  bound  to  protein  in  the  blood  serum  of 
the  ox. 

Nucleoprotein  is  found  in  the  blood  plasma  and  the  serum  as 
well  as  in  the  corpuscles ;  of  leucocytes  it  forms  the  chief  constitu- 
ent. (See  Plosz,  1871;  A.  Kossel,  1881,  1893;  Halliburton  and 
Friend,  1889 ;  Halliburton,  1895 ;  Lilienf eld,  1894;  Pekelharing,  1895; 
Bang,  1903,  1904;  and  Liebermeister,  1906). 


150  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

.  The  amount  of  phosphocarnic  acid  was  studied  by  Panella 
(1902c).  He  found  an  average  of  0.3631  percent  in  the  blood  of 
dogs,  0.282  percent  in  the  blood  of  rabbits  and  0.0674  percent  in  the 
blood  of  calves. 

The  most  frequently  quoted  early  analyses  of  the  blood  of  dif- 
ferent species,  including  human  subjects  in  different  conditions, 
are  those  of  Becquerel  and  Rodier  (1844),  Jiidell  (1868),  von  Bunge 
(1876),  Jarisch  (1877)  and  Miroczkowski  (1878). 

We  have  selected  from  the  data  of  a  number  of  authors  items 
showing  the  amounts  in  the  blood,  of  the  different  phosphorus  com- 
pounds, and  of  a  few  other  constituents  which  may  be  considered 
in  connection  with  the  same.  Some  of  these  are  given  in  connection 
with  studies  on  the  changes  brought  about  in  pathological  condi- 
tions, and  are  included  in  the  tables  with  such  pathological  data.  The 
following  table  of  Abderhalden's  is  quite  unusual  with  regard  to 
the  number  of  species  considered  and  the  completeness  and  care 
with  which  the  analyses  were  made. 

Abderhalden  has  tabulated  these  analyses  to  show  the  amounts 
of  the  individual  constituents  contained  in  1000  parts  by  weight  of 
blood,  in  1000  parts  of  serum  and  in  1000  parts  of  corpuscles.  He 
calls  attention  to  the  similarity  of  composition  of  the  serum  from 
the  different  animals,  and  the  great  similarities  of  composition  of 
the  blood  of  two  animals  of  the  same  species.  It  is  also  very  evi- 
dent that  the  bloods  of  the  three  kinds  of  ruminants  (cattle,  sheep 
and  goat)  agree  much  more  closely  with  each  other  than  with  that 
of  the  carnivora  (dog  and  cat)  or  with  that  of  the  horse,  pig  and 
rabbit.  "The  ruminants  show  a  considerably  lower  phosphoric  acid 
content  than  the  carnivora  or  the  horse,  pig  and  rabbit.  Perhaps 
in  the  ruminants  there  is  an  organic  acid  present  with  the  phospho- 
ric acid." 


PHOSPHORUS  METABOLISM 


151 


=?  4  2 o<j§ g 

^£^3"®      oti1... 

.  «  P*.  o 

'b- 


»a«9Q»PSs.»s^_  £5*8;  »S  £2 


fcj-  09 


E, 


©©©•     ©CO- 


mmco 

(f.OS-3 
CO  CO  CO  CO 

coco  bo  co 
en     to© 


©  ©to©©;  ©to© 

©  Hobb'  i-'*-b 

tO  Mrf^-COOs!  OlOH 

CO  MOOCOOO.  COMtO 

OS  If.        COCO  CO 


©M©O©00.  pOO 

p.  m  bo  to  bs  to  •  en  'as 

^oowoooi'  if-os 

00  if.CnMlf. 


op©;  ©m; 

bo  if.  b  •  bs  to  • 

coos  to;  Mlf." 

Ol©CO.  OnOs 


-q  tO- 
to  OS) 

to  00. 


mmco 

CO  CO  CO 

(£.co  oscn 
oibn°co<i 
en      —a  ifa- 


©copo;  ©too 

J-'i^bb-  "m^o 

OlCOCO-q-  O300  M 

moooo  to.  COCOcn 


OOMOM»' 


-q  CO  OiCO  OS 


©©©.     ©©MO 


MMCO 

CO  If.  OS 
MO-      If.  COlf.  CO 


Os  Or  o  •  OSPHO 
OSOl  tO-  if.  Mlf.|f. 
OS  MCOt      M00COO1 


o  to  pp.  ©to© 

ptooo;  m bi b 

i^oocoos.  co to o 

C0lf.COO3-  coos© 


co.  v^-  rf=- 

©pM©t-*Oi.  cn<) 

-qoso'^-bb'  bos 

CO  CO  o  CO  00  if.  •  Cnlf. 

cocoosOif.     • 


©       pOp-  ©©MO 

bbbt  -a  m  too 

w     -aoco-  oi-'osif». 

oo      cococo:  osif^cncn 


MMCO 
lf^OS-3 

MO.    top'en-q 
O  en '.    co  Os  m  -q 

MOT     OS       ©M 

-aos; 


©     ©to©o.    ©coo     ooooo^.    coif. 


MCOOO 

coo  to  OS 

©OlOlMl      lf.C0cO 


MCOo       OSCOcOCO-qO-     00  CO 

|C.lOO       CDM-qOsCOOr     co© 

C0if.-3Oscn 


OS|f.O-      OS  M  M  O 

coonco;    tDM-3  os 
-qcnoi.    if.  to>f.co 


co  on 
en  on 

If*  OS 


CO  if.  CD  CO  00  M  •      COM 
oo  if.  -3  co  os  os ;     Ol-q 
00C0CO       COOSMCOO 


MMCO 

.      M  MCO  M 

©©©•      ©O©©        •     •      MO-      00C0O1M 


toono- 

enco-q  o 

•      CO  OS- 

-aonooco 

i^mm; 

if.cocnco     ; 

;   coo; 

OS        OSOl 

COlfs.  If.. 

-3OSO100      • 

.     COM 

©CO©©;  ©CO©  ppt-ippp- 

m  if*  b  b  •  m  bo  b  co  b  m  b  bo  b  • 

oioto-a;  ostoM  to  >f.  to  to  to  os ; 

if.  co  os  oo.  o.tf-M  coo-^ooco 


OsGQ 


C3 


-*3 


2 

**£  o 


tog 


2 

M  o 


©3 


to  as  o 


oo3 


f 

co 

M 

CO 

>    >TJ 

8  M 

^  M 

s»   LJ 

Hi!  W 
8  f 
3  o 

00  O 
^§ 
*2 

!»  fO 
_      HH 

M  02 

©   CO 

O    »fl 

o  9 

u_i  |— i 
3  H 

©    GO 

o    _ 
&-  O 


tr* 
co 


152 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


_  & 


<B    P* 


2     Mo 

B    cog 


>    g- 


a    m 


m   i 


p!p£,b-S  <=  2  _  P*£2.2  B\S  S  »  °  » 


o°9 


£,s°  B  o 

'"  g  So 
"S      o  P'.—.p'tr' 


hi  i-s 


O.   .ffl 


«■£-     p  5 

p  2 

5 


tO  Ol  ©  •  if-  tO  ©  © 
Ol  ©  O  ■  tD  tO  Ol  tO 
tO  ©  Ol :   tO  00  tf»-  M 


O  -1  •  O  ID  O  to 
to  to  •  CO   ©  en 

O1C0. 


O       0(000       ©10©       OOM©©©- 


HWOO.     M©© 

©-a  tooo  •    -cioo© 
O^OOHl    -q  if>.-q 


to  -q  Oi  to  Ol 


©  ©©..   ©©©© 


tooio-  Oi  to-q  o 
tO-q©-  lf>-CO©  to 
GO  Ul  ©.     Ol  ©  ©  CO 


MM-      tO  tO  OOO 


00  00  00  CO 
tO^-  1>0(© 

too. 


©to©©     ©to© 


HOiOO-  H(DO 
©  M  tO  Qo;  -qHM 
CO©-q©I     tO -3  © 


}H  00  ©  00  -J  Ol  •  ©Hi 
©Oloo©©©-  CO© 
O'O  00  MOO 


o©o.   ©o©o 


to©coto 

MM.     O  CO  tO  tO 
tO  M  •      00  M  00  © 

too-     toOOiCn 

©ol   to 


o  too©-    ©  too 


MlHO©-  MOO 

©OOCOOO"  -qoo 

(H  OOOO:  MOO  00 

©©©Ol  tDtH  to 


wOHOO00 
P*  ©  M  00  V]  tO  ' 


©©©•   ©op© 

to  ©  O  •     Ol  to  bo  © 


otopp.  ptoo 

!_i  t-  ©  o :  m  bo  © 

oioi  to<i  •  -a-q© 

©CO-qCOl  rf^COOO 


OMHOOOl. 


)£>■  CO  10  Ol  ©  £-  •      ©© 
©  Ol  rf^  ©  -q  M  *      ©CO 

if»--4>f*©©     ; 


©to 
bo  © 


MtOCO 

co©o  to 
©©If**"- 


M  tO- 

©  ©: 

Ol©. 


©  pMOO;  ©t-3© 

©  M  -1  ©  ©  ■  MOO 

CO  MO!  to  en*  tooo© 

CO  tO  tO  M  tO  I  CO  Ol  © 

©  O0C0MCO  -J  CO  If. 


"ooppp.    ©ih 

P-  ©  GO  M  Ol  ©  '      ©  tO 
OM©MOltO-      -5C0 

f*HcooH    : 

CO  © 


©OO-     ©M- 


MMbS 

to  tOOl  1^- 

©  •      M  ©  •     O  Ol  CO  CO 


OlCO. 

o 

O- 

©  to- 

O00 CO  00 

©to- 

en 

ti' 

co  ©; 

Ol       lf»  -3 

toco! 

© 

tt-- 

i—  -'• 

O  ©top©;  pt-SO 

0  m  to  b  ©  •  M  ©  © 
^  iH©to©;  oito© 

01  C"  M  S  ©  •  to  if".  tO 


C0Ol©CO00©.      MM 
©  O  Cn  M  ©  Ol  '     Ol  if* 

cotOMCo-a     : 


«5P 


-»9 


2 

.JO  °" 
»d  o 
Mg  p. 

On 
too'n 


©cc 

oo2 


cog 


03  e  o 
t°g  P" 


wg 


2 

,«2 


-*P 


pp 

cog 


«S? 


fr- 
>■ 

Hi 

GO 

H 
CO 

O 
*J 

a 

H 

CLfH 

re  o 
s-O 
S.O 
^^ 
2  O 


co  <1 

•"0O 

3d 

oH 
oG0 

tdO 
o 


GO 

o 
o 

o 

f 

o 

H 

O 


PHOSPHORUS  METABOLISM 


153 


Weiske  (1886)  analyzed  the  blood  of  lambs  in  connection  with 
a  study  of  the  effects  of  ingestion  of  acids.  From  this  work  the  fol- 
lowing data  are  quoted. 

.   INORGANIC  ELEMENTS  OF  LAMB'S  BLOOD 
Weiske  (1886)  Percent,  Dry  Basis 


Animal 

P 

N 

K2O 

Na20 

CaO 

MgO 

Feed,  6  months 

0.150 
0.150 
0.130 

15.255 
14.960 
15.090 

0.415 
0.430 
0.305 

2.220 
2.445 
0.395 

0.066 
0.074 
0.065 

0.029 
Trace 
Trace 

Normal  hay,  barley- 
Hay  treated  with   di- 
lute H2SO4,  barley 
Killed  at  beginning1  of 
the  experiment 

Lamb  III 

For  comparison  with  the  data  on  blood  of  mammalia  we  quote 
the  following  figures  for  constituents  of  the  blood  of  birds,  from 
the  work  of  Weiske. 

DETERMINATIONS  ON  THE  BLOOD  OF  BIRDS 
Weiske  (1889)  Percent,  Dry  Basis 


Bird 

Mineral  matter 

N 

CaO 

Mg-O 

P2O5 

5.99 
6.14 
5.57 
5.52 
5.85 
5.27 

14.84 
15.19 
15.22 
15.37 
15.11 
15.98 

0.12 

0.08 
0.06 
0.07 
0.10 
0.04 

0.08 
0.06 
0.06 
0.07 
0.07 
0.09 

1.69 
1.71 
1.82 
1.55 
1.79 
1.77 

Fowl 

Fowl 

Fowl 

Bergell  (1898b)  gives  the  following  values  of  the  phosphorus 
content  of  the  blood  ash  of  cattle :  Blood  of  calf,  8.36,  6.73,  7.83  per- 
cent P205;  of  cow,  6.21  percent;  of  ox,  4.17,  5.06,  4.64,  5.66  per- 
cent. He  thinks  there  is  a  significance  in  the  ratio  of  serum  phos- 
phorus to  erythrocyte  phosphorus. 

Burger  and  Beumer  (1913b)  found  lecithin  in  the  blood  cells  of 
man  and  sheep  in  minute  amounts  only.  The  phosphatids  consist 
chiefly  of  sphingomyelin,  with  some  cephalin,  an  ether-soluble  diam- 
inomonophosphatid,  and  a  water-soluble  phosphatid. 

PHOSPHORUS  COMPOUNDS  IN  MILK 

GENERAL  COMPOSITION 

It  is  the  idea  of  Duclaux  (1884a,  1884b,  1884c,  1893,  1894)  that 
in  milk  both  casein  and  calcium  phosphate  exist  partly  in  suspen- 
sion and  partly  in  solution,  the  casein  being,  in  fact,  an  equilibrium 

Note:_  For  a  general  discussion  of  the  constituents,  properties  and  changes  of  composi- 
tion of  milk  the  reader  is  referred  to  the  review  and  summary  of  literature  by  Raudnita 
(1903);  also  to  Basch   (1903)   and  Kastle  and  Roberts    (1909). 


154 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


of  the  three  conditions  solid,  colloidal  and  dissolved.  The  phos- 
phates in  suspension  are  thought  to  be  those  of  calcium,  of  magne- 
sium and  of  iron  or  aluminum,  the  calcium  phosphate  in  suspension 
being  about  twice  the  amount  of  that  in  solution. 

Vaudin  (1894a,  1894b,  1895, 1897b)  finds  alkali  citrates  in  milk, 
which,  aided  by  the  presence  of  lactose,  he  considers  essential  for 
holding  the  calcium  phosphate  in  solution. 

Richmond  (1901)  looked  upon  casein  as  being  a  compound 
containing  sodium,  calcium  and  phosphorus  united  in  the  milk  with 
Ca3  (P04)  2,  the  salts  of  the  milk  including  also  both  mono-  and  di- 
basic phosphates.  Richmond  considers  that  of  the  0.75  percent  of 
ash  in  milk  0.27  percent  is  salts  combined  with  casein,  the  remaining 
0.48  percent  being  salts  in  solution  in  the  milk  plasma. 

Soldner  (1888)  thought  that  the  calcium  united  with  casein  is 
not  a  phosphate  but  the  oxide  only. 

Barille  (1909a,  1909b),  from  microscopical  and  chemical  exam- 
inations of  milk,  has  put  forth  the  idea  that  in  it  there  are  crystal- 
lizable  compounds  of  the  phosphates  with  carbonic  acid  which  he 
calls  "carbonophosphates,"  and  which  he  considers  beneficial  for  the 
assimilation  of  milk.  Pasteurization,  he  states,  precipitates  and 
decomposes  these  carbonophosphates  almost  completely  and  hence  is 
injurious.  Sterilization  with  ultraviolet  rays  leaves  the  carbono- 
phosphates intact,  and  is  recommended.  The  data  may  be  of  in- 
terest. 


CARBONOPHOSPHATES  IN  MILK  AS  AFFECTED  BY  STERILIZATION 

Barille  (1909)— Grams 


Sample 
No. 

Phosphoric 

acid  per  liter,  as  P2O5 

Carbonic  acid  per  liter 

Milk  analyzed 

Insoluble 
phosphates 

Soluble 

phosphates 

Carbono- 
phosphates 

In  free  state 
and  as  car- 
bonophos- 
phates 

As  carbon- 
ates and 
dissociated 
carbonates 

From  Department  at 

1 
2 
3 
4 

5 

5+ 

6 
6+ 

7 

7+ 

0.812 
0.728 
0.767 
0.750 

0.521 
0.690 

0.666 

0.882 

0.660 
0.710 

0.677 
0.519 
0.711 
0.640 

0.540 
0.630 

0.530 
0.598 

0.350 

0.370 

0.214 
0.045 
0.011 
0.050 

0.372 
0.079 

0.372 
none 

0.210 

0.190 

0.052 
0.046 
0.043 
0.064 

0.078 
0.068 

0.132 
0.075 

0.091 

0.080 

0.136 

Commercial  pasteurized         ( 

Prom  dairy  at  Paris 

Same  pasteurized  by  us. .. 
From  dairy  near  Paris 

Drawn  in  our  presence 

Same  pasteurized  by  us. . . 
From  dairy  in  Paris 

0.076 
0.072 
0.077 

0.140 
0.065 

0.173 
0.042 

0.148 

Same  submitted  to  ultra- 

0.107 

PHOSPHORUS  METABOLISM 


155 


Aside  from  inorganic  phosphates  and  casein,  in  whatever  com- 
binations they  may  be  present,  there  are  other  phosphorus  com- 
pounds reported ;  a  phosphatid,  usually  considered  to  be  a  lecithin,  a 
nucleon  similar  to  the  phosphocarnic  acid  of  muscle,  and  perhaps  an- 
other phosphorized  protein,  the  opalisin  of  Wroblewski  (1894a, 
1894b,  1898).  He  found  opalisin  present  in  abundance  in  human 
milk,  in  less  quantity  in  mare's  milk,  and  in  very  small  amount  in 
cow's  milk. 

The  relative  amounts  of  the  constituents  in  milk  are  not 
constant,  as  is  shown  by  the  data  we  submit,  and  the  milks  of  dif- 
ferent species  of  mammalia  differ.  Naturally,  a  part,  but  not  all, 
of  the  variations  appearing  in  the  tables  are  due  to  the  use  of  differ- 
ent methods  of  determination.  Schmidt-Mulheim  (1883)  compares 
the  first  and  last  portions  drawn  at  milking.  The  last  is  always 
considerably  richer  in  fat,  but  the  fat-free  serum  does  not  vary 
much  in  composition. 

The  table  of  Ellenberger,  Seeliger  and  Klimmer  (1902)  compar- 
ing the  composition  of  the  milk  of  woman,  cow  and  ass  will  serve  to 
show  the  general  composition  of  milk,  and  some  of  the  variations 
with  species.  Ellenberger's  special  study  was  on  the  milk  of  the 
ass.      (See  also  Ellenberger,  1899, 1902.) 


COMPOSITION  OF  ASS'S,  WOMAN'S  AND  COW'S  MILK 
Ellenberger,  Seeliger  and  Klimmer  (1902) — Percent 


Ass's  milk 

Woman's  milk 

Cow's  milk 

Dry  matter   .... 

Albumin    and 
globulin      .... 

Fat     

91.23 
8.77 
1.50 
0.94 

0.53 

0.1 
1.15 

0.02 

6.0 

0.4 

86.4 

13.6 

1.6 

1.0 

0.5 

0.12 

4.8 

0.06 

6.6 

0.25 

88.0 

12.0 

3.3 

3.0 

0.3 

0.06 

3.5 

Lecithin     

Milk  sugar    .... 
Salts     

0.05 
4.5 
0  75 

We  quote  below  the  averages  of  Katayama's  (1908)  extensive 
analyses  of  the  milk  of  different  pure  breeds  of  cattle,  and  of  the 
milk  of  unimproved  native  cattle  from  different  countries.  (The 
averaging  has  been  done  in  the  main  by  the  compilers). 


156 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


AVERAGE  COMPOSITION  OF  MILK  OF  VARIOUS  BREEDS  OF  CATTLE 
Katayama  (1908) — Percent 


Num- 

ber 

Dry  sub- 

Pat 

Nitrogen 

Ash 

Lime 

Phosphor- 

Breed 

of 

sam- 
ples 

stance 

CaO 

ic  acid 
PaOs 

Silesian  red   .... 

3 

13.39 

4.63 

0.580 

0.705 

0.152 

0.216 

Red-brown  East 

2 

14.14 

5.65 

0.677 

0.842 

0.193 

0.222 

Scheinf  elder 

3 

13.03 

4.57 

0.519 

0.703 

0.152 

0.212 

Voigtlander    .... 

3 

12.39 

4.19 

0.543 

0.778 

0.162 

0.209 

Simmenthal    .... 

3 

13.94 

4.66 

0.625 

0.781 

0.196 

0.238 

Black-spotted 

East    Friesian 

3 

12.03 

3.22 

0.605 

0.784 

0.193 

0.247 

3 

12.86 

4.32 

0.545 

0.738 

0.166 

0.236 

Wilstermarscher 

3 

12.37 

3.87 

0.524 

0.699 

0.155 

0.200 

Swiso     .  , 

3 

12.45 

3.98 

0.574 

0.739 

0.164 

0.219 

East  Prussian 

6 

11.12 

3.16 

0.449 

0.713 

0.154 

0.188 

Average  of  these 

improved  breeds  .. 

12.55 

4.08 

0.547 

0.742 

0.167 

0.216 

10.76 

2.02 

0.429 

0.666 

0.143 

0.180 

14.95 

6.36 
3.36 

0.682 

0.863 

0.227 

0.273 

3 

12.28 

0.510 

0.747 

0.185 

0.227 

2 

13.23 

4.94 

0.489 

0.654 

0.170 

0.200 

2 

10.44 

0.75 

0.516 

0.732 

0.200 

0.280 

Dutch  East  Africa 

6 

12.40 

4.08 

0.680 

0.708 

0.176 

0.225 

Buffalo    

1 

9.69 

4.37 

0.336 

0.467 

0.150 

0.153 

Average  of  native 

12.04 

3.60 

0.567 

0.695 

0.178 

0.225 

9.69 

0.70 

0.336 

0.467 

0.150 

0.153 

13.60 

5.89 

0.649 

0.795 

0.204 

0.296 

DISTRIBUTION  OF  PHOSPHORUS  IN  MILK 

As  to  the  distribution  of  phosphorus  among  the  constituents  of 
milk  we  do  not  find  close  agreement  in  the  evidence. 

Raudnitz  (1903)  gives  the  following  figures  for  phosphorus  in 
various  conditions,  which  must  be  looked  upon  as  approximations. 
They  differ  considerably  from  other  estimations  which  we  quote. 

PARTITION  OF  PHOSPHORUS  OF  MILK— (Raudnitz,  1903) 


Total  P2O5 
Percent 

Casein  P2O5 

Lecithin 

P2O5 
Percent 

Preformed  phosphoric  acid 

Authors 
mentioned 

Species 

Percent 

Percent 
of  total 

Percent 

Percent 
of  total 

0.24 

0.047 

0.135 

0.0581 

0.012 

0.0117 

about  21 
25 
8.2 

0.005 
0.006 
0.005 

0.18 
0.034 

rest 

about  79 

72 
nearly  90 

Baginsky 

ers 

Schlossmann  (1905b)  compared  different  milks,  and  the  follow- 
ing values  show  a  contrast  between  human  and  bovine  milk  in  the 
amount  of  phosphorus  present  in  inorganic  form. 


PHOSPHORUS  METABOLISM 


157 


PARTITION  OF  PHOSPHORUS  IN  MILK— (Schlossmann,  1905b) 

Total 

P2O5 

Gm.  per 

liter 

Casein  P2O5 

Lecithin  P2O5 

Nucleon  P2O5 

Rest,  as 
inorganic 

Species 

Gm.  per 
liter 

Percent 
of  total 

Gm.  per 
liter 

Percent 
of  total 

Gm.  per 
liter 

Percent 
of  total 

Percent 
of  total 

Cow 

,1.65 
0.455 

0.58 
0.16 

35 
35 

0.091 
0.16 

5.5 

35 

0.09 
0.153 

5.5 
33 

54 

0 

Marre  (1911)  finds  nearly  all  of  the  lecithin  of  cow's  milk  in 
buttermilk.  He  states  that  pasteurization  of  milk  decomposes  the 
lecithin. 

Forbes,  Beegle  and  Mensching  (1913)  found  56  percent  of  the 
phosphorus  of  skimmed  cow's  milk  to  be  inorganic. 

The  partition  of  phosphorus  as  given  by  Gilbert  and  Posternak 
(1903, 1905)  is  as  follows : 

GRAMS  P205  PER  LITER  OF  MILK 


Species 

Total 

In  casein 

In  lecithin 

In  nucleon 

Total  organic 

Woman      .... 

1.81 
0.43-0.47 

0.580 
0.132 

0.091 
0.153 

0.087 
0.171 

0.758 
0.456 

Stoklasa  (1897)  gives  the  average  lecithin  P205  of  cow's  milk 
as  0.091  gm.  per  liter,  or  5  percent  of  the  total,  and  that  of  woman's 
milk  0.153  gm.  per  liter,  or  35  percent  of  the  total,  which  agrees 
with  Schlossmann.  Jensen's  (1904,  1905a,  1905b,  1905-6)  details, 
reported  from  15  milk  analyses,  distinguishing  between  the  phos- 
phorus of  casein  and  that  of  insoluble  or  soluble  salts,  make  the 
casein  phosphorus  about  25  percent  of  the  total  in  cow's  milk. 

Values  reported  by  Paton,  Dunlop  and  Aitchison  (1899-1900) 
indicate  that  in  goat's  milk  the  phosphorus  of  casein  and  lecithin 
is  about  29  percent  of  the  total ;  that  of  nucleon  is  about  16  percent, 
and  the  inorganic  about  55  percent  of  the  total. 

Sikes  (1906)  made  about  three  hundred  estimations  of  phos- 
phorus, and  a  somewhat  smaller  number  of  determinations  of  cal- 
cium, on  human  milk.      The  following  are  his  conclusions : 

Phosphorus:  1.  The  average  amount  of  P205  in  human  milk 
during  the  first  fortnight  after  the  commencement  of  lactation  is 
0.0297  percent,  the  variations  being  between  0.0140  and  0.0522  per- 
cent. 


158  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

2.  The  percentage  of  the  non-proteid  P205  averages  0.0169, 
and  of  the  proteid  P205  0.0124,  for  the  total  number  of  cases.  For 
primipara  and  for  multipara  the  numbers  are  0.0160  and  0.0129,  and 
0.0183  and  0.0116,  respectively. 

3.  The  ratio  of  the  proteid  P205  to  the  total  P205  averages 
42.3  percent,  but  it  varies  between  14.7  and  77  percent.  In  primip- 
ara the  average  is  44.6  and  in  multipara  38.8  percent. 

4.  The  chart  for  primipara  shows  that  the  total  amount  of 
P205  increases  up  to  the  ninth  day,  while  that  for  multipara  shows 
a  maximum  a  little  earlier.  After  this  in  both  the  P205  falls  to  a 
lower  level. 

5.  The  variations  mentioned  are  chiefly  due  to  the  non-proteid 

PA. 

6.  The  proteid  P2Og  does  not  vary  much  on  successive  days. 
This  uniformity  is  rather  more  noticeable  in  multipara  than  in 
primipara. 

7.  If  one  assumes  that  the  average  amount  of  proteid  in 
human  milk  is  2  percent,  the  numbers  mentioned  will  give  the  av- 
erage amount  of  P205  in  it  as  0.62  percent.  For  primipara  the 
number  will  be  0.64  and  for  multipara  0.58  percent. 

Calcium.  1.  The  average  amount  of  calcium  in  human  milk 
in  the  first  fortnight  after  delivery  is  0.0301  percent. 

2.  In  primipara  the  average  is  slightly  lower  than  in  multip- 
ara. 

3.  The  major  part  of  the  calcium  is  combined  with  the  proteid. 
The  ratio  of  the  proteid  calcium  to  the  total  calcium  is,  on  the  aver- 
age, 84  to  100. 

4.  The  variations  of  the  calcium  from  the  average  stated  are 
small. 

5.  The  curve  giving  the  percentages  on  successive  days  after 
delivery  is  more  uniform  than  the  similar  curve  of  the  percentages 
of  P20D  in  the  same  samples. 

6.  When  both  the  calcium  and  the  proteid  were  estimated  on 
the  same  samples,  the  calcium  was  found  to  average  1.06  percent  of 
the  total  proteid. 

Njegovan  (1913)  investigated  the  lecithin  content  of  milk  by  a 
carefully  considered  method,  and  concluded  that  the  existence  of 
lecithin  in  milk  is  doubtful. 

MINERAL  CONSTITUENTS  OF  MILK 

We  have  brought  together  from  various  sources  a  number  of 
individual  or  average  determinations  of  the  mineral  constituents  of 
the  milks  of  different  species,  grouping  them  as  to  species.  No  at- 
tempt was  made  to  consider  the  methods  of  analysis. 


PHOSPHORUS  METABOLISM 


159 


1 

ercent.     Percent  of  ash.      Mean  of  2 
d  by  compilers. 

er  and  Soldner,  1900  and  1902,1903. 
lactation  period, 
tation  period. 

X 

CO 

p 

s 

xl 

en 

3 
3 

s 

CO 

0) 

>. 

c4 

3 

d 
_o 

(4 

s 

1-1 

CD 

3 
o 
+J 
u 

CU 
Pi 

a 

o 

o 

1 

■3 

i 

co 

cu 
p 
o 

to 
u 

'S 

o 
u 

b» 

X 

•d 

41 

3 

a 
.2 

"S 

0 
o 
u 

■d 

4  1 

a   § 

CO        CN 

0> 

0) 

~2       t» 

O              d  "rf        ho 
et           »°       d 

£           3S-Crt 
&         g.25  2 
o         £„  XI  ca 

CO              J«,|g 

5      atl° 

d          o  9  <4   . 

■2    A     3 

g  .-s  1 

.  o      a 
rtx!      S  „5fJ 

ti.             O  ai   ra 

°^  sz* 

3  „     "d  Sxi 

en  rt      -p  k,,—. 
14  CU        3  &"d 

en 

.2 
+3 
d 

'S  . 

a  2 

83 

a  IN 

p  B  52 

i  o  d 

tut 

X 
•d 
cu 

s"i 

cu  P 

p 

ao 

8.x 

^OT»< 

"dri  o 

CU        +j 

5  fc!  S 

en 

s 
"S 

a 

o 
0 

>. 

X 

•d 

cu 
p 
3 

p 

s 

o 
o 
CM 

"3 

3 
rt 

1 

a 

a 

d     I 

~«     xi 
•d     -p 
d  „:o 

« °  s 

cu  c" 

O             O  O,  >.  _  5     .        o 

i»  a       >,2-3l§8.0 
fed        ^  m  c  rt  feo  S>2 

S3        «?» °S     8- 

SiH             +j  btT=|+J'd   u  °  o 
P  w            ft  3  s  D  o  "■«  3 

S       o5    &    (3    § 

£5£dEs 

^•5  o 

8 

IN 

d 

1 

rt.  a  rt  u  El 
w  9  O  c4  , 
II  8  0W  o 

0  ~        i< 

01  <       (J 

S53 

go 

Kg 

o 
en 

3cd""    . 

5  8)° 

Z    o 

2dg 
°^-§ 
si 

cuO! 

r-ICO 

t-       CM 

_, 

COON 

O""* 

i*l  t—  0CO3C35IO 

CO  CO  CD 

in 

THOO 

y— v        .—1          TH 

OT       CO 

CM^^COlOOi        ,— s 

COCM^H 

CO 

y-si*i  C7i  CO  ^H  Cv  in 

CDHO 

r-H 

O 

THCD 

o     r-     co 

lO      th 

TTHCOCOIOCD        CO 

CD  COO 

o 

CM        OS  CO  CM  CO  O  OS  CO 
1— 1        t-  i—l  i-H  CM  i—l  CO  i—l 

t^OCM 

oo 

t-     o     o 

o     o 

oooo^-i     io 

OCMi— 1 

OHH 

Jh 

1HH 

oo 

OT       d       O 

d     o" 

ooooo     <a5 

ooo 

o 

d     Nocidddo' 

odd 

d 

do 

CO 

CO 

1 

i-it- 

"-1     SB 

o~* 

•         OSCM-* 

1— IT*ICO 

cooomooco 

CMCOf* 

in 

o 

t~r- 

^N       CM        OO 

r~     co     cm 

>-iot;-h 

^tOOCOh.       ^*» 

CM  00  00 

o 

IO       /— ^CM  CM  *—<  Tt<  0O  CD 

TMin 

CO  CM  CO 

CMCOCOIOC35        ^H 

000102 

I» 

1—1        O^OOJGOOOO} 

ooo 
coS 

& 

oo 

(MOO 

ooo 

O^Hr-ICM-H       i-H 

iHCOH 

i-H 

CM        lOTtlCMCOCMCMCn 

COtHCO 

TH 

do 

co     o     o 

odd 

OOOOO       TH 
CM 

ooo 

o 

d     ddddddd 

odd 

d 

do 

CO 

o 

inoo 

CM           jH 

CM 

lococo     ^-, 

C35-J5     • 

TH 

co         r^^mcocoo 

'  CMIO    . 

CD 

oo 

^«.    o     o 

CO        i-H 

1-I.HT-lrHO         1—1 

om    • 

rH 

CO              CO  TH  i— 1  C0 1— 1  CM 

oo 

OT        O        O 

CO       o 

OOOOO        i-H 

OO      • 

o 

O              OOOOOO 

oo    " 

CO 

oo 

I-H        O        O 

o     o 

ooooo 
OOOOO      S- 

oo    • 

o 

CO              OOOOOO 

oo    ; 

o 

oo 

oo 

i-H        O        O 

d     d 

oo    • 

o 

o         daod^iss 

do   • 

d 

do 

o 

bo 

in  CM 

"*       CM 

CO       CO 

CM 

O        lOHO 

IC5COCO 

CO  00  CO  TM  i—l  CM 

COOT 
OO 

<-»    io     m 
ley    o     o 

in     co 
o     o 

IOCOCNtJHtH        ^^, 
COrM-HCMCM          Ttt 

CDCor^ 

t— ICM^H 

N 

OS       C—^O  TM  CD  in  TH  in 

i-H     t^  in  .—1  co  ^  cm  in 

COOTCO 

CO 

t^Oi 

OO 

CD       O       O 

o     o 

ooooo     »o 

OOO 

o 

CO       CDOOOOOO 

ooo 

o 

oo 

OO 

CM       O       O 

d     d 

OOOOO       CM 

o'oo 

o 

d     fModdddd 

odd 

d 

oo 

inm 

SP     ^! 

co-*  SI 

CM 

00        CDCMO) 

T*l  1-1 1—1 

t^-COTHTHr^TH 

t-IOOT 

CO 

o 

i4 
O 

coi— i 

^-**    r~     oo 

t^COCD 

COCDCO^hCO       y-N 

CM^pICO 

o 

00      /^hiooi-.h3 

m« 

Ttl        CO        CO 

CO  CO  CO 

COOCMIOIO        UO 

in  co  oo 

-H 

CD        CD  l>-  TH  i— 1  05  TH  OS 

OO 

th     o     o 

ooo 

OHi-lNH        1*1 

r-ICMiH 

i—l        t— CMCMCMi— ICMOO 

CMtHtH 

TH 

COTH 

o'o 

CO      o      o 

ooo 

OOOOO       i-H 

ooo 

d 

d     THoddddd 

odd 

d 

do 

» — 

- — 

o 

TdCO 

oo     co 

t-OS— J 

O       05100 

rHi-HCO 

OOTHCOt^OO 

THO 

/~\    TH     1— 

mo-** 

CMCOCOlOi— 1        /«*» 

i— IIOCO 

o 

00        ^-^t^  CD  CM  i—l  CO  0O 

NO 

O       TH       rH 

cocot^- 

COCOi— c*i— 1        1*1 

«»■* 

t^ 

CD        Oi  CO  QO  in  CD  1^  03 

r-r^co 

rt 

OO 

05      o      o 

ooo 

COOOOi-H        Oi 

OiHO 

o 

O       l>t— OOOOi— 1 

ooo 

o 

i-HO 

8 

oo 

eo     d     d 

t— i 

ooo 

OOOOO       o" 

o'o'o 

d 

d     icidoo<ssi<s 

odd 

d 

o'o' 

»-ICM 

00        TH 

THOOT 

CM       lOCOCD 

1 

ooco 

TH  C--  in  CM  tH  CO 

cd  co  in 

00 

O 

THTH 

/— >       O        CO 

ooooco 

corjiTjuoca     ^^, 

0OCMCO 

in 

i— 1       ^t^  CD  TH  CO  in  ^H 

t^to 

S    S    8 

COTJIO 

NCCOON        *— 1 

CM'HOO 

r- 

t*.     cd  — ^  o  co  co  r-  in 

S3 

M 

oo 

ooo 

OOHHH         T*1 

1-ICNrW 

1—* 

1— 1        1— 1 1— 1 0  CM  rt  O  CM 

ooo 

rH 

oo* 

§  ®  °" 

odd 

OOOOO       K5 

ooo" 

d 

o     ooodddd 

odd 

d 

do 

xi 

CO 

o 

cS     So 

occo 

CMOT 

oco-*co     /-n 

1 

00  OS 

CD        COOT 
*■ s       CO        iHt> 

co 

CO 

00 

Sgo 

CO 

< 

CM  CO 

O        OT 
CO        »-H 

OH 

)   i— IT*ICO       o 

co^it~oo     ex 

ISO) 

r~ 

TH        TH        t—  t— 
I>       CO       ISt^ 

OOOO 

o 

CO 

CO  CM 
rtCO 

do 

O        O 

oo 

OOOO       CO 

oo 

d 

g    d    do 

i-H 

dr-ii-* 

rH 

i-i  r-5 

8 
a  o 

o 
o 

d 

(4 

*d     tj  -d  d 

•d 

■d 

thOC> 

O        CM        CM 

NOn 

hcmtxoj'Ji     in 

tH        i— 1 00  OT  00  OT  05  03 

Oi 

TH 

t-iH 

O       O       o 

O^^H 

i— lor^-cct--     oo 

OJO^COCOOO       oo 

CO     CO 

i— 1        0O  OT  Cn  OT  OT  OT  OT 

r-- 

h  <4 

00     - 

Oi     OS 

OT        00  O0  00  CC  GO  00  0O 

CO 

CO 

ccoo 

OT 
00 

rH        1-1        r-i 

»-!  1— 1 »— 1  T— 1 1— i        1— i 

1-1        Hi- li— 1  i—l  i—l  i-H  i-i 

f-11— 1 

O 

x 
p 

3 

IS 

x    x    xt 

"3 

i-  3  "5 

<U        jj         d 

Is  §1 9*8  a 

»      CU 

u 

o 

■  .   .       a       3  3  3 

TJ-J          CU          CU    CU   CU 

SSfjfeS  53222 

a       PX  rtX  14  «J  ca 

Call)  U  n   U  rjrj  rt 

x  -r  a)  nM  -a  .^  -d  T3  -d 
1/1  >  W  k  x  W  X  X  X 

g 

2  «p 
5  >  ° 

■S«  a 

cu  fj  d 

cu 

ho 

d 

41 

■d 

41  -3 
be  d 

S  S  b 

a  u^ 

°-9 

3     en 

£     3 

&  1-1 

u 

X 

n 

3 
3 

n 

X 

n 

in    in    a 

o 

<i 

<J 

to 
41 

8 

Pi 
K 

3  d"  P 

d     d*     d 

d  d 

d 

o 

•  'S 
:  tti-p 

P  d"2 

o  3  d 

He 

3  3  = 
WWE 

sic 

3        3        = 

W    W    P 

14  c4 

EH 

3  3 

WW 

1" 

3  u 

1 

3 

3  c 
ML 

i 

c 

" 

1 

I 

c 

c 
L 

s 

c 

p      p  p  , 
CU        CU  cu  +■ 
cu       cu  cu  n 

X      XX  c 

en    cnwC 

s 

t 

s 

s 

fa 

c 

c 

1 

b 

c 

c 

t  ho 
o 
a 

160  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

On  the  basis  of  analyses  of  milk  and  of  its  serum  and  its  ash, 
Soldner  (1888)  gives  the  following  as  the  forms  and  amounts  of 
mineral  constituents  present  in  one  liter  of  milk:  NaCl  0.962,  KC1 
0.830,  KH2P04 1.156,  K2HP04  0.835,  potassium  citrate  0.495,  dimag- 
nesium  phosphate  0.336,  magnesium  citrate  0.367,  dicalcium  phos- 
phate 0.671,  tricalcium  phosphate  0.806,  calcium  citrate  2.133,  cal- 
cium oxide  of  casein  0.465.  He  considered  only  calcium  oxide  to  be 
united  with  the  casein. 

Vaudin  (1897a)  made  determinations  of  total  mineral  matter, 
and  of  earth  phosphates  in  milk  of  different  breeds  of  cows,  from 
different  countries,with  different  feeds,  and  when  different  quanti- 
ties of  milk  were  being  produced,  with  the  conclusion  that  none  of 
these  factors  has  much  influence  on  the  quantity  of  ash  or  on  its 
earth  phosphate  content.  The  total  mineral  matter  found  ranges 
from  7.05  to  7.83  gms.  per  liter  in  all  the  normal  cases  except  two 
(two  are  8.10  and  8.66  respectively)  and  the  earth  phosphates  from 
3.30  to  4.10  in  all  cases. 

Friedenthal  (1911)  has  computed  the  number  of  ions  of  the  in- 
organic constituents  present  in  the  milk  of  several  species,  especially 
the  number  supplied  by  diluted  cow's  milk  and  by  woman's  milk. 
These  computations  are  based  on  analyses  by  Schloss.  A  table 
gives  the  anions  and  cations  (K,Na,Mg,Ca,Cl,P04  and  the  rest  as 
citric  acid)  per  liter  of  the  milk  of  each  species.  The  P04  values 
stated  are  :  human  milk  2.73X10'3,  cow's  milk  13.46X103,  sow's 
milk  16.8X10-3,  dog's  milk  35.76X10"3,  rabbit's  milk  70.18X10"3, 
ass's  milk  15.7X10"3.  The  relation  of  these  numbers  to  one  another 
indicates  the  relative  amounts  of  inorganic  phosphate  in  the  milks, 
showing  human  milk  to  have  a  low  value,  and  that  of  the  dog  and 
rabbit  very  high  values  in  this  respect. 

Schloss  (1911)  reports  the  following  analyses  of  woman's  milk 
from  24-hour  samples,  the  first  being  a  mean  of  analyses  of  milk 
from  8  women,  the  second  and  third  representing  the  mixed  milk  of 
16  and  15  women  respectively. 

ANALYSES  OF  WOMAN'S  MILK— Parts  per  Liter 

Mean  of  8  analyses        Mixed  milk  I  Mixed    milk  II 

Fat     37.88  40.225  35.87 

Nitrogen      1.847  1.904  1.897 

Total  ash    1.839  1.913  1.838 

CaO      0.3758  0.3856  0.380 

MgO     0.0857  0.0761  0.0725 

£ia2°       0.1886  0.1623  0.1958 

£20      0.5291  0.5292  0.5360 

*8°6     0.4046  0.4469  0.3829 

CASEIN  CONTENT  OF  MILK 

For  each  of  the  organic  phosphorus  compounds,  also,  we  have 
collected  a  table  from  the  individual  determinations. 


PHOSPHORUS  METABOLISM 


161 


I 

ho 

-: 

tfl                                                                                      g 

i 

3                                                                                      >> 

.•a             8- 

CS                                                                                     «w 

ft                   ho 

-P                                                                    .  ho 

I-. 
o 

a         e 

w                                           +J  a 

co             h                                                         x^  S 

X 

to 

8             %. 

an  of  8.    2Colostrum.    3Mean  of 

per  lOOcc.     Mean  of  8.     Some  fi 
last  drawn. 

n  of  2. 

uding-  traces  of  globulin. 

n  of  2.    Quoted  by  Schlossmann, 

ted  by  Schlossmann. 

rage  up  to  time  of  doubling-  weig 
rage  after  doubling-  weigfht  of  yoi 
h  salts  in  combination, 
n  3rd  to  116th  day  of  lactation. 

3 

0} 

>>                w 

to 

4) 

f 

V} 

o 

X 
•M 

0 

■a 

o 

X 

o 

a 

' 

ent  proportions  ol  hay  and  he 

er  lOOcc. 

er lOOcc 

uted  from  26. 

ige  of  many  analyses. 

a;  computed  b 
's  tables. 
's  tables, 
compilers, 
by  compilers, 
of  6  monthly 

o 

CO 

15;  2  method: 
d  from  Konig 
d  from  Konig 
ad  from  40  by 
35;  computed 

100  cc.  Mean 

n 

■s 

.■as  3  °  » 

(4 
X 
u 

V 

a 

is  e^e-S 

a  ft  ft  ft  a  n 

V                ri                   ra                       viraO              I?  P  ±>  a 

<§       a       s          e^i  3      >  >r  s 

ff           O             S                 w§0)         <«!£& 

s  a  a  a  <«  n- 

200020 

.s 

"55 

to 

CD     in     50                      i-it~o,H 
en     en     cm                      t~i— ii>-oj 

en  cm 

Tft 

men 

CM 

rt 

CO       CM       CO                              COOOTflTti 

coo 

iA 

to 
u 
o 

XI 
-p 
O 

O 

to 

•d 

a 

M 

Buffalo 
Elephant 
Gamoose 

Cat 

Rabbit 

Guinea  pig 

Guinea  pig 

M^5 

2 

C3 

ho 
O 

<N       (MOJ 

CD       OOTti 

0 

Tfl        TflTfl 

he 

oo  r» 

m 

lOi— t 

Tf( 

to 
o 

s 

coco 

t>5 

CO 

Tf( 

'o 

4-> 

CO                                                                                                          r-{CO 

CM 

en 

S 
P. 
CO 

O)                                                                                                          C3SIO 

-■ 

CM 

cog 

3 

co                                                                  cm"cm 

CM 

tS"" 

CM 

rH  <M 

en 

p. 

TMt~-                                                                                             00t- 

CD 

CMCO                                                                                                      OO 

>?cn          co 

XI 
CO 

iar-5                                                                ■*•* 

gTf!          co- 
co 

,— ICN1 

to 

00  CO 

-f 

Tff 

l> 

to 

CnOOt- 

CO 

00 

CD 

<J 

oo'o 

o 

o 

o 

O 

a 

O                                                                                                                   t-H 

CO 

rt 

1-5  CD                                                                                                              _• 

CO 

a 

3 

w 

0.7- 
1.04 

1.2 

1.810-C 

o 

CO 
CD 

o 

0.18-1 
1.03 

0.49 

en 

en 

CM 

1 

OO                           lO              O                                                                             CO 

ooco  Tti   co     en 

CO                           CM.             IO                           O                                              CO 

O 

OOIO     CD     Tfl        CD 

O^OOO 

O 

co                  cm          co                  co                               co 

CO 

cmcm   cm   cm     cm 

CO^COCM 

£  ".2 

cdX)                                                              rt  nj'd'd 

J2 

a 

COOOCMCOCO        00OO        O        Tflt--              CO  CO  CO  CO  r-l  ,— ( 

CM 

CMcoTfiioiOior^oo 

JH  o  -y 

t^COOOOOGOQO        0OC0CO        CO        COCO              OJCiQOJOO 
0O0OOO0O0O00        000000        00        00  00  C  P- 00  00  00  00  C»  CO 

o 

oooooooo 

OOOOHH 

<u  CJ5 

CD 

cocococococncocn 

«  ^ 

-V- 

Meigs,  A.  V 

Schmidt-Miilheim. . 

X 

s  • 

:  .  :   :  .  :  •- 

:  :  :i3A 

o 

XI 

a 
■4 

X 

1= 

a 

.1 

> 

< 

.5 

A 

1 

ft 

f 

c 

o 

s 

■d 

c 

— -c 

ftc 

^1    • 
o    • 

is  : 

.."a 
ea 

C  <« 

-  gs,. 
§JsS 

W    coy 

1 

bderhalden 

bderhalden 

bderhalden.... 
bderhalden 

llenberger.    See 
grer  and  Klimm 
Drdas  and  de 

S 
to 

is 

o 

o  2 

rt  £ 

I 

I 

on  Szontagh 

on  Szontagh 

stertag-andZun 
anSlyke  and  Bo 

worth 

;ach — extremes 
iach — mean 

igel  and  Frehn 

1 

5 

> 

s 

e 

c 

fc 

as 

<<«(£ 

WW 

M 

E- 

t>t>WC!> 

^MWSI 

162 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


Rogers  (1909)  gives  the  casein  content  of  buttermilk  as  2.4,  of 
kefir  2.56-2.59  and  of  kumiss  0.77-0.85  percent. 

Engel  and  Frehn  (1910),  computing  the  casein  content  of  human 
milk  from  the  nitrogen  loss  of  the  milk  on  precipitation  with  acetic 
acid,  report  the  examination  of  31  samples  from  several  wet  nurses, 
at  different  times  in  the  lactation  period,  and  of  four  samples  of 
mixed  milk,  from  which  they  conclude :  "The  relative  casein  value  of 
human  milk  undergoes  wide  variations,  even  with  the  same  individ- 
ual, for  which  no  laws  could  be  recognized.  The  factors  which 
have  empirically  been  taken  to  be  of  great  influence  for  the  compo- 
sition of  human  milk,  like  the  time  of  lactation,  individuality,  the 
phase  of  the  emptying  of  the  breast,  or  the  length  of  intervals  be- 
tween nursing,  play  no  recognizable  part  in  determining  the  casein 
content." 

PHOSPHATID  CONTENT  OF  MILK 

The  content  of  phosphatid  in  milk  varies  with  the  time  of  lac- 
tation, and  is  reduced  by  heating,  as  well  as  by  skimming.  Fetzer 
(1910)  has  noticed  that  the  milk  from  animals  suffering  from  mas- 
titis contains  less  lecithin  than  that  from  healthy  animals.  It  was 
further  noted  that  where  a  diminution  in  the  lecithin  content  took 
place  there  was  a  corresponding  decrease  in  the  fat  content. 

Very  few  of  the  studies  have  distinguished  lecithin  in  milk  from 
other  phosphatids  possibly  present.  W.  Koch  (1906)  thought  he 
identified  both  lecithin  and  cephalin  in  milk. 

Wrampelmeyer  (1892)  reports  that  100  gm.  of  butter  contains 
0.0451  gm.  P205,  indicating  0.017  percent  lecithin. 

PHOSPHATID  OF  MILK  OF  DIFFERENT  SPECIES  AS  REPORTED  BY 
VARIOUS  AUTHORS— Reported  as  Percent  Lecithin 


Date 

Species 

Author 

Cow 

Human 

Ass 

Dog 

Sheep 

Goat 

Mare 

Notes 

1897 

1900 
1902 

J-1908-j 

1909b 

1909 

0.090- 

0.113 

0.054 

0.05 

0.0364- 

0.1163 

0.0765 

0.0629 

07) 
0.0449 

0.170- 
0.186 
0.058 
0.06 
0.0240- 
0.0799 
0.1329 
0.0499 
(10) 

0.024' 

0.0058- 
0.0393 

0.0165 
(6) 

0.17 

0.0509- 
0.1672 

0.0833 
(4) 

0.0349- 
0.0753 

0.0488 
(ID 

0.0073- 
0.0174 

0.0109 
(8) 

Gm.  per  100  ccmilk 

Nerking  and 

Extremes 

Average  and  no.  of 

Kida 

analyses 

Mean  of  4 

NUCLEON  CONTENT  OF  MILK 


The  nucleon  content  of  the  milk  of  woman,  the  goat  and  the  ass 
are  shown  by  the  following  figures  to  be  somewhat  nearly  the  same, 
while  cow's  milk  seems  to  contain  only  half  as  much  of  this  constit- 
uent. 


PHOSPHORUS  METABOLISM" 


163 


NUCLEON  CONTENT  OF  MILK  OF  DIFFERENT  SPECIES  AS  REPORTED 
BY  VARIOUS  AUTHORS— Computed  as  Carnic  Acid,  Percent  or  Grams 

per  100  c.  c.  Milk 


Date 

Species 

Author 

Cow 

Human 

Ass 

Goat 

Notes 

1896 

1896 

1897 
1897 

1902 
1905, 
1908 

0.0583 

0.0502 
0.0566 

0.124 
0.1315 

0.120 
0.1 

0.110 

by  compilers. 
Gm.  per  100  cc.  Decreases  on  standing. 

Ellenberger,  Seeligrer 
andKlimmer 

Average  for  year.  Mean  of  3  computed 
by  the  compilers. 

Blumenthal  (1896)  found  that  the  0.5021  gm.  carnic  acid 
(measure  of  phosphocarnic  acid)  in  a  liter  of  cow's  milk  decreased  to 
0.2430  gm.  while  the  milk  stood  at  room  temperature  for  eight  days. 
This  is  taken  as  indicating  that  phosphocarnic  acid  is  a  source  of 
the  decomposition  products  of  milk. 

SOME   COMPARISONS   OF  WHOLE  MILK,   SKIMMED  MILK  AND   CREAM 

A  considerable  part  of  the  lecithin  of  milk  and  a  smaller  pro- 
portion of  the  casein  are  removed  with  the  cream ;  hence  skimmed 
milk  is  richer  in  casein  and  much  poorer  in  lecithin  than  whole  milk. 
Bordas  and  de  Raczkowski  (1902b),  however,  claim  that  the  lecithin 
is  not  wholly  removed  in  the  cream,  while  Glikin  (1909b)  does  not 
agree  with  them,  and  says  that  if  the  cream  is  completely  separated 
by  centrifuging  the  lecithin  is  also  completely  removed.  Following 
are  Bordas  and  deRaczkowski's  analyses  of  milk,  3.200  liters  of 
which  gave  2.800  liters  of  skimmed  milk  and  0.370  liters  of  cream. 


PHOSPHORUS  COMPOUNDS  IN  WHOLE  MILK,  SKIMMED  MILK  AND 

CREAM— Grams  per  100 


Whole  milk 

Skimmed  milk 

Cream 

2.88 

0.176 

0.0044 

0.0124 

0.058 

3.24 

0.184 
0.0013 
0.0037 
0.018 

1.15 

0.096 

0.0252 

0.0691 

0.334 

The  figures  below  are  from  d'Hont  (1890),  who  obtained  12 
liters  of  skimmed  milk  and  3.5  liters  of  cream  from  15.5  liters  of 
whole  milk. 


164  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  6 

ANALYSES  OF  WHOLE  MILK,  SKIMMED  MILK  AND  CREAM 

Percent 


Whole  milk 

Skimmed  milk 

Cream 

Ash    

5.05 
4.70 
3.50 
0.79 
0.22 
0.226 

0.025 

5.05 

3.62 

0.788 

0.214 

0.22 

21.95 
3.32 
3.02* 
0.585 

0.155 
0.17 

*  Prom  Kjeldahl  nitrogen   determination. 


Dornic  and  Daire  (1910)  report  that  buttermilk  contains  more 
lecithin  than  whole  cow's  milk,  and  more  even  than  woman's  milk. 


CHANGES  OF  PHOSPHORUS  COMPOUNDS  AS  A  RESULT  OF  HEATING  0E  STANDING 

Baginsky  (1883)  found  that  heating  milk  to  120°,  and  de- 
hydrating as  in  Scherff' s  method  of  preservation,  did  not  decrease 
the  ratio  of  phosphorus  in  casein  to  phosphorus  in  the  whey. 

Bordas  and  de  Raczkowski  (1903)  noted  that  milk  heated  to 
various  temperatures  over  the  free  flame,  on  the  water  bath,  or  in 
the  autoclave  loses  through  decomposition  a  certain  amount  of  its 
lecithin.  Diffloth  (1904)  also  studied  the  effects  of  heating  on 
milk.  He  found  that  a  like  effect  is  produced  by  simply  allowing 
the  milk  to  stand.  Comparisons  were  made  of  the  phosphorus 
changes  due  to  standing  and  to  heating  as  in  sterilization  or  pas- 
teurization. Total  phosphorus  was  determined  on  the  solids  taken 
up  by  acetic  acid,  soluble  inorganic  phosphorus  on  the  clear  liquid 
left  after  complete  coagulation,  soluble  organic  on  the  same  (this  is 
interpreted  as  lecithin) ,  and  the  insoluble  phosphorus  by  difference. 
According  to  these  figures,  and  the  author's  interpretation  of  them, 
lecithin  decomposition  takes  place  even  when  the  milk  stands  at  or- 
dinary temperature,  but  heating  causes  a  much  greater  change.  The 
duration  of  heating  seems  to  have  a  greater  influence  than  the  de- 
gree of  heat ;  but  with  the  same  length  of  time  of  heating  the  dim- 
inution of  assimilable  phosphates  (formation  of  insoluble  phos- 
phates and  decomposition  of  lecithin)  increases  as  the  temperature 
increases.       (See  table  on  next  page.) 

Kida  (1909)  reports  the  following  percentages  of  reduction  in 
the  lecithin  content  of  cow's  milk  as  a  result  of  heating  for  thirty 
minutes  at  different  temperatures:  80°,  7.52  percent;  80°,  11.39  per- 
cent; 95°,  25.27  percent;  100°,  21.22  percent. 


PHOSPHORUS  METABOLISM 


165 


Grosser  (1913)  finds  that  boiling  milk  5-30  minutes  does  not  al- 
ter the  freezing  point  or  the  phosphorus  content  of  that  portion  of 
the  milk  which  passes  through  a  Bechold  filter — colloids  are  held 
back. 

PHOSPHORUS  PARTITION  IN  COW'S  MILK  AS  AFFECTED  BY  STAND- 
ING AT  15°  C.  AND  AT  HIGHER  TEMPERATURES 
Diffloth  (1904)— Grams  P2Os  per  Liter  of  Milk 


Time  of  standing1 

Temper- 
ature 

Total 
phos- 
phorus 

Soluble 
inorganic 

Soluble 
organic 

Insoluble 

Increase 
in  insol- 
uble 

Decrease 
in  leci- 
thin 

15°C 

60° 
60° 
95° 
95° 
110° 

4.58 
4.58 
4.58 
4.58 
4.58 
4.58 
4.58 
4.58 

1.92 
1.92 
1.96 
1.85 
1.81 
1.82 
1.79 
1.75 

2.12 

2.08 
2.01 
1.90 
1.60 
1.50 
1.47 
1.38 

0.54 

0.58 

0.61 

0.83   i 

1.17 

1.26 

1.32 

1.45 

o'.64 
0.07 
0.29 
0.63 
0.72 
0.78 
0.91 

0.04 

0.11 

0.22 

0.52 

0.62 

0.65 

0.74 

CHANGES  IN  MILK  WITH  PROGRESS  OF  LACTATION 

Cow's  Milk.  Houdet  (1894)  made  a  chemical  study  of  colos- 
trum of  the  cow.  From  his  conclusions  we  quote  the  following: 
"The  composition  of  colostrum  remains  about  the  same  from  the 
time  the  liquid  appears  until  after  the  birth  of  the  young.  In  the 
days  following  the  birth  marked  changes  occur  at  once."  Among 
the  changes  noted  is  this,  "The  calcium  phosphate  and  other  salts, 
abundant  at  first  in  both  soluble  and  insoluble  condition,  diminish 
gradually  until  about  the  fourth  or  fifth  day,  when  they  show  about 
the  same  content  as  in  normal  milk." 

In  the  main,  the  observations  of  Trunz  (1903)  correspond  with 
this  last  statement  of  Houdet,  though  perhaps  there  was  longer 
time  in  getting  back  to  normal,  (6-7  days  for  one  cow).  Trunz 
made  complete  analyses  of  the  milk  of  two  cows  at  frequent  inter- 
vals (30  samples  for  each)  throughout  lactation.  We  have  com- 
puted a  few  summary  statements  from  the  phosphorus  data.  For 
further  detail  the  original  article  should  be  consulted. 

PHOSPHORUS  OF  MILK  AS  AFFECTED  BY  PERIOD  OF  LACTATION 
Trunz  (1903)— Grams  per  1000 


Day  of  highest 
phosphorus  content 

First  4  days 

Average  of  later  values 

Cow 
No. 

Total  P2O5 

P2O5  in  salts 

Number 

of 
samples 

Total 
P2O5 

P2O5  in 

Mini- 
mum 

Maxi- 
mum 

Mini- 
mum 

Maxi- 
mum 

salts 

655 
674 

Day  of  calving1 
Day  after  calving- 

2.227 
2.137 

3.284 
3.144 

1.534 
1.564 

2.564 
2.557 

25 
19 

2.015 

2.075 

1.529 
1.622 

166 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


According  to  the  statements  of  Kort  (1899),  based  on  a  number 
of  investigations,  the  content  of  cow's  milk  in  mineral  matter,  par- 
ticularly in  phosphoric  acid  and  calcium,  decreases  more  or  less  reg- 
ularly as  the  time  of  birth  of  the  calf  draws  near,  even  in  spite  of 
decreased  flow  of  milk ;  the  colostrum  is  rich  in  ash ;  the  percentage 
of  phosphoric  acid  and  calcium  in  the  milk  is  lowest  when  lactation 
is  at  full  height,  though  the  absolute  quantity  is  then  the  greatest. 

Schrodt  and  Hansen  (1885)  determined  the  ash  composition  of 
the  milk  of  the  same  cows  (10)  at  seven  dates  between  January 
and  September,  the  times  of  calving  having  been  October  15  to  Jan- 
uary 1.  The  cows  were  in  stall  from  January  to  May,  and  in  pas- 
ture from  June  to  September.  The  phosphoric  anhydride  data,  in 
percent  of  the  ash,  are  as  follows:  January  1,  23.11;  March  30, 
23.11 ;  May  20,  24.61 ;  June  16,  22.41 ;  July  26,  23.59 ;  August  30, 
26.51 ;  and  September  26,  25.41. 

Bordas  and  de  Raczkowski  (1902a)  report  the  following  figures, 
and  say  that  both  the  phosphoric  acid  and  the  lecithin  constantly 
fall  off  in  amount  after  the  time  of  calving,  animals  of  the  same 
breed,  and  fed  in  the  same  way,  showing  the  milk  of  the  first  month 
to  be  highest  in  lecithin. 

PHOSPHORUS   CONSTITUENTS   IN  MILK   OF   COWS  AT  DIFFERENT 
STAGES  OF  LACTATION  (Bordas  and  de  Raczkowski,  1902)— Percent 


Breed 

Jersey 

Norman- 
dy 

Date  of  calving 

Cow  with 
calf 

July  4 

July   1 

June    12 

May   13 

Not  with 
calf 

Casein   content 
Total  phosphoric 

Other  organic  phos- 

As  glycerophos- 

3.86 

0.218 

0.0049 

0.0139 

0.0654 

3.17 

0.204 

0.0068 

0.0198 
0.0909 

2.90 

0.200 

0.0044 

0.0124 
0.0582 

2.89 

0.164 

0.0033 

0.0100 
0.0472 

3.04 

0.168 

.    0.0033 

0.0100 

0.0472 

,  2.30 

0.148 

0.0041 

0.0116 
0.0545 

2.61 

0.156 

0.0031 

0.0093 
0.0436 

Note:   The  date  of  taking  the  samples  is  not  given;   samples  probably  taken  all  on  the 
same  date. 


The  general  question  of  the  influence  of  the  stage  of  lactation 
on  the  composition  and  properties  of  milk  has  been  studied  by  Eckles 
and  Shaw  (1913).  They  analyzed  samples  taken  throughout  the 
year  from  11  cows  of  four  breeds  which  were  kept  on  a  uniform 
ration  of  alfalfa  and  grain,  and  from  5  other  Jersey  cows  not  kept 
on  uniform  rations.  The  casein  seldom  went  beyond  the  limits  of 
80  to  82  percent  of  the  total  protein,  and  averaged  81,4  percent. 
Both  total  protein  and  casein  were     abnormally    high    following 


PHOSPHOEUS  METABOLISM 


167 


parturition,  then  declined  continuously  till  the  third  or  fourth  week, 
when  the  minimum  was  reached.  They  then  remained  fairly  con- 
stant until  near  the  end  of  the  lactation  period,  when  they  rose  rap- 
idly and  reached  the  maximum  at  the  end  of  the  period. 

Human  Milk.  Schlossmann  (1905b)  gives  the  following  values 
for  the  total  phosphorus  in  woman's  milk  at  different  times,  indicat- 
ing no  characteristic  change  with  duration  of  lactation. 


12 

21 

213 

191 
(right 
breast) 

191 
(left 
breast) 

191 

(right 
breast) 

191 

(left 
breast) 

Grams  per  P2Os  per  liter.  . 

0.65 

0.56 

0.52 

0.37 

0.35 

0.51 

0.34 

Engel  and  Frehn's  opinion  that  the  wide  variations  in  casein 
content  observed  in  human  milk  are  not  due  to  the  stage  of  lactation 
has  been  mentioned  above  (see  p.  162).  Edlefsen  (1901)  gives  a 
table  of  the  gross  analysis  of  human  milk  at  different  periods.  Case- 
in appears  regularly  to  decrease  with  advance  in  period  of  lactation. 

ANALYSES  OF  HUMAN  MILK  AT  DIFFERENT  PERIODS  OF  LACTATION 
Edlefsen,  (1901)— Percent 


Day  of  lacta- 
tion 

Total  protein 

Casein 

Albumin 

Tat 

Sugar 

3     

12     

48      

103     

116     

2.695 

1.875 

1.00 

0.843 

0.835 

1.810 

1.160 

0.44 

0.375 

0.310 

0.885 
0.715 
0.560 
0.468 
0.525 

3.225 
3.035 
3.64 
3.415 

4.105 

3.59 
5.15 

7.06 

5.835 

5.95 

Valenti  (1905,  1908)  found  that  the  content  of  nucleon  in  hu- 
man milk  is  greatest  during  the  first  and  second  months  of  lacta- 
tion, generally  decreasing  from  the  third  to  the  sixth  month,  in- 
creasing again  during  the  seventh  month,  and  then  remaining  con- 
stant until  the  twelfth  month. 

RELATIONS  OF  THE  COMPOSITION  OF  MILK  TO  THAT  OF  THE  YOUNG 

VonBunge  (1874),  in  connection  with  studies  of  the  alkalis  of 
milk  and  other  articles  of  diet  and  of  the  young  organism,  intro- 
duced the  idea  which  became  known  as  Bunge's  law  that  the  compo- 
sition of  the  ash  of  milk  (with  the  exception  of  the  iron)  corre- 
sponds closely  with  that  of  the  ash  of  the  total  organism  of  suck- 
lings. VonBunge  (1886,  1889)  and  his  pupils  Abderhalden 
(1899b,  1899c)  and  Proescher  continued  to  investigate  the  question, 
considering  first  whether  the  ash  of  blood  and  blood  serum  also  have 


168 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


the  same  composition,  and  whether  there  are  characteristic  differ- 
ences between  the  species  in  this  respect  running  parallel  in  the  milk 
and  the  body  of  the  young.  Pages  (1894)  criticized  the  supporters 
of  Bunge's  law  on  the  ground  that  it  was  too  sweeping,  and  that 
it  was  applied  to  all  ash  constituents,  not  distinguishing  those 
which  are  essential  for  the  growth  of  the  bodies.  We  have  not  seen 
Pages's  thesis.  A  little  later  Proescher  (1898)  and  Abderhalden 
(1899a,  1899d)  compared  the  composition  of  the  milk  of  each  spe- 
cies in  relation  to  the  time  required  by  the  young  of  that  species 
to  double  their  live-weight.  The  numerical  support  for  the  idea  of 
such  a  relation  is  best  illustrated  by  the  table  below,  taken  from 
Abderhalden  (1899d,  p.  594).  Protein,  ash,  lime  and  phosphoric 
acid  in  the  milk  of  this  series  of  animals  all  increase  in  the  same 
order  as  does  the  rate  at  which  the  young  double  their  weight. 

RELATION  OF  THE   COMPOSITION   OF   THE   MILK   OF   DIFFERENT 

SPECIES  TO  THE  TIME  REQUIRED  FOR  THE  YOUNG  OF 

THE  SAME  SPECIES  TO  DOUBLE  IN  WEIGHT 

Abderhalden  (1899) 


Species 

Time  required  for  the 

new-born  animals  to 

double  the  body  weight 

Days 

100  parts  by  weight  of  milk  contain 

Protein 

Ash 

Lime 

Phosphoric  acid 

180 
60 
47 
22 
15 
14 

m 

9 
6 

1.6 
2.0 
3.5 
3.67 
4.88 
5.21 
7.00 
7.44 
10.38 

0.2 

0.4 

0.7 

0.7713 

0.8406 

0.8071 

1.02 

1.3282 

2.4998 

0.0328 

0.124 

0.160 

0.1974 

0.2453 

0.2489 

6.'  4545 
0.8914 

0.0473 
0.131 

Cattle 

0  197 

Goat 

0  2840 

0  2928 

0  3078 

Cat 

Dog- 

0.5078 

Rabbit 

0.9967 

With  regard  to  human  milk,  and  the  body  of  the  human  infant, 
investigation  has  yielded  results  which  do  not  support  the  theory, 
or  which  at  least  show  that  the  parallelism  is  less  close  here  than 
for  other  animals.  See  the  work  of  Cornelia  deLange  (1897,  1900), 
of  Hugounenq  (1899b,  1900),  and  of  Camerer  and  Soldner  (W.  Cam- 
erer,  Jun.  1900,  1902a;  W.  Camerer,  Jun.  and  Soldner,  1900,  1903; 
Soldner,  1902). 

Another  suggestion  has  been  made  by  Burow  (1900)  to  the  ef- 
fect that  there  may  be  a  significant  relation  between  the  lecithin 
content  of  the  milk  of  different  species  and  the  brain  weight  of  the 
suckling  young  of  the  same  species.  For  cattle,  dogs  and  men  it 
was  found  to  hold  that  the  greater  the  relative  brain  weight,  the 
higher  is  the  lecithin  content  of  the  milk,  reckoned  in  percent  of  the 
protein.  "Here  also  nature  has  matched  the  needs  of  the  sucklings 
with  the  composition  of  the  milk." 


PHOSPHORUS  METABOLISM 


169 


COMPARISON  OF  THE  RELATIVE  LECITHIN  CONTENT  OF  MILK  AND 

THE  RELATIVE  BRAIN  WEIGHT  OF  THE  YOUNG  OF 

DIFFERENT  SPECIES— (Burow,  1900) 


Mean  composition  of  milk 

Species 

Lecithin 
Percent 

Protein 
Percent 

Ash 
Percent 

Lecithin:  pro- 
tein=X:100 

Brain  weight:  body  ■weight 

Cattle 

0.051 

0.17 

0.058 

3.84 
8.05 
1.90 

0.69 
1.00 
0.24 

X=1.40 
2.11 
3.05 

1:370  (mean  of  4,  7-8  wks.  old.) 

STUDIES  OF  THE  PHOSPHORUS  OF  EGGS 

General  Composition  and  Phosphorus  Content.  All  of  the  types 
of  phosphorus  compounds  found  in  eggs,  with  the  exception  of  nu- 
clein,  were  noted  by  Gobley  (1846,  1847)  in  his  researches  on  hen's 
eggs  and  on  the  eggs  of  carp  (1850a).  The  compounds  are  nuclein, 
phosphorized  fat,  nucleoalbumin  and  traces  of  inorganic  phosphate. 

Liebermann  (1888a)  made  chemical  examinations  of  different 
portions  of  the  yolk  and  of  the  white.  He  also  followed  the  special 
chemistry  of  the  embryo  through  to  the  maturity  of  the  chick.  He 
found  nuclein  and  lecithin  in  the  germ;  but  no  phosphorus  in  the 
ether  extract  of  the  yolk ;  he  concluded,  therefore,  that  the  lecithin 
present  is  not  free,  but  is  in  chemical  combination,  perhaps  with  vi- 
tellin,  as  previously  suggested  by  Hoppe-Seyler.  Extraction  with 
water,  or  with  dilute  acid  failed  to  reveal  any  phosphate  directly 
recognizable  without  ashing. 

According  to  Kaas  (1906),  the  amount  of  phosphorus  in  the 
white  is  increased  by  remaining  in  contact  with  the  yolk.  T.  B.  Os- 
borne and  Campbell  (1900c)  report  0.122  percent  phosphorus  in  the 
ovalbumin  of  egg  white. 

Carl  Voit  (1877a)  gives  the  average  phosphoric  acid  content  of 
the  white  of  one  egg  as  0.007043  gm.,  which  would  be  0.197  percent, 
and  that  of  the  yolk  as  0.20386  gm.,  which  would  be  2.57  percent. 
Lebbin  (1900,  1901)  found  an  average  of  0.22  percent  P205  in  the 
whites  of  hen's  eggs  and  1.43  percent  in  the  yolks,  or  0.636  percent 
in  the  whole  edible  part.  Forbes,  Beegle  and  Mensching  (1913) 
found  only  traces  of  inorganic  phosphorus  in  the  edible  portion  of 
the  hen's  egg. 

According  to  Hammarsten  (1911,  p.  600,  604)  Poleck  and 
Weber  found  1000  parts  of  the  ash  of  the  white  of  the  egg  276.6- 
284.5  parts  potash,  235.6-329.3  soda,  17.4-29  lime,  17-31.7  magnesia, 
4.4-5.5  iron  oxide,  238.4-285.6  chlorine,  31.6-48.3  phosphoric  acid 
(P205),  13.2-26.3  sulphuric  acid,  2.8-20.4  silicic  acid  and  96.7-116 


170 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


parts  carbon  dioxide.  In  1000  parts  of  the  ash  of  the  yolk  were  found 
51.2-65.7  parts  soda,  80.5-89.3  potash,  122.1-132.8  lime,  20.7-21.1 
magnesia,  11.90-14.5  iron  oxide,  638.1-667.0  phosphoric  acid,  and 
5.5-14.0  parts  silicic  acid. 

Carpiaux  (1908)  gives  the  makeup  of  the  ash  of  a  fresh  egg 
as  being  17.37  percent  K20,  22.87  percent  Na20,  10.91  percent  CaO, 
1.14  percent  MgO,  0.39  percent  Fe203,  37.62  percent  P205,  0.32  per- 
cent S03  and  8.98  percent  CI. 

Malcolm  (1902)  has  given  us  a  comparison  of  the  chemical 
composition  of  the  yolks  of  eggs  laid  by  different  hens  of  the  same 
breed,  and  on  the  same  diet ;  also  of  the  different  eggs  laid  by  the 
same  hen  on  a  constant  diet.  His  tables  are  given  in  part  below. 
The  conclusions  are:  1.  The  percentage  of  lecithin  in  egg-yolk 
varies  considerably.  2.  The  percentages  of  protein,  fat  and  phos- 
phorus in  the  yolks  of  eggs  from  the  same  hen  are  in  very  close 
agreement,  while  there  are  very  considerable  differences  in  eggs 
from  a  number  of  hens,  even  where  the  breed  is  the  same. 


ANALYSES  OF  EGG- YOLKS— (Malcolm,  1902)  Percent 

Egg- 
No. 

Fat 

Nitrogen 

P2O5 

Ether  extract 

Chloroform  extract 

Percent 

P2O5 

content 

Percent 

P2O5 
content 

Laid  by   hens   all   of 
one  breed,  and  fed  J 
on  maize  and  barley  ) 

Laid  by  one  hen. .          -j 

1 
2 
3 

4 
5 
6 
7 
8 
9 
10 
11 
12 
13 
14 
15 
16 
17 
18 

27.786 
29.453 
28.501 
28.662 
31.887 
28.956 
29.114 
30.080 

30.' 124 
29.565 

3L043 
31.575 
30.431 
30.120 

36! 265 

3.004 
2.620 
2.648 
2.674 
2.676 
3.123 
2.877 
2.674 
2.605 
2.822 
2.846 
2.762 
2.575 
2.552 
2.607 
2.754 
2.797 
2.747 

1.489 

L356 
1.527 
1.396 
1.517 
1.511 
1.569 

i.*47i 
1.654 
1.543 
1.392 
1.425 
1.441 
1.477 

L506 

28.662 

28.956 
29.114 
30.080 

0.1408 

0.2120 
0.1884 
0.1838 

31.418 

31.423 
32.659 
33.100 

0.2085 

0.2419 
0.1930 
0.2036 

THE  NXTCLEOALBUMINS  OF  EGGS 

The  nucleoalbumin  of  hen's  eggs,  ovovitellin,  seems  to  be  pres- 
ent in  chemical  combination  with  lecithin,  and,  as  isolated  and  pre- 
pared, it  usually  contains  lecithin.  According  to  T.  B.  Osborne  and 
Campbell  (1900b),  it  is  extracted  as  a  mixture  of  several  protein- 
lecithin  compounds  which  may  be  called  lecithin-nucleo-vitellins. 
Plimmer  (1908)  reports  another  protein  containing  less  phosphorus 
than  vitellin. 

The  corresponding  compound  from  fish  eggs,  called  ichthulin, 
was  especially  studied  by  Valenciennes  and  Fremy  (1854)  in  their 
general  consideration  of  the  composition  of  salmon  eggs;  but  G. 
Walter   (1891)   gave  it  a  more  particular  study,  as  he  obtained 


PHOSPHORUS  METABOLISM  171 

it  from  the  eggs  of  carp,  and  he  decided  that  vitellin  and  ichthulin 
may  be  considered  identical.  Hammarsten  (1905a)  isolated  a  nu- 
cleoalbumin  from  the  eggs  of  perch  which  yielded  a  much  larger 
proportion  of  pseudonuclein,  on  pepsin  digestion,  than  Walter  had 
found  from  the  ichthulin  of  carp  eggs. 

NUCLEIC  ACID  01*  FISH  EGGS 

J.  A.  Mandel  and  Levene  (1906b)  obtained  a  nucleic  acid  from 
unfertilized  fish  eggs  which  in  its  chemical  makeup  resembled  nu- 
cleic acids  of  plant  origin  more  than  those  from  animal  substances 
generally.  Helene  Tschernorutzky  (1912a)  found  indication  of 
about  1.2  gm.  nucleic  acid  in  100  gm.  of  dry,  ripe,  unfertilized  her- 
ring eggs  which  had  been  extracted  with  alcohol  and  ether.  The 
phosphorus  content  of  the  substances  not  removed  by  alcohol  and 
ether  was  not  all  accounted  for  by  this  nucleic  acid. 

THE  PHOSPHATIDS  OF  EGGS 

A  large  part  of  the  study  of  phosphatids,  under  the  name  of 
lecithin,  has  been  made  on  that  obtained  from  eggs.  (See  Diaco- 
now,  1867a,  1867b,  1868b;  Strecker,  1868;  Bergell,  1900;  Cousin, 
1903 ;  Henriques  and  Hansen,  1903 ;  Laves,  1903a,  1903b ;  Wintgen 
and  Keller,  1906;  Erlandsen,  1907;  Stern  and  Thierf elder,  1907; 
Frankel  and  Bolaffio,  1908;  MacLean,  1908b,  1909b,  1909c;  Serono 
and  Palozzi,  1911;  Riedel,  1912;  Trier,  1912,  1913b.) 

Armand  Manasse's  (1906)  determinations  of  the  amount  of 
lecithin  obtainable  from  egg-yolk  by  different  treatments  show  re- 
sults nearly  enough  alike  (range,  8.856-9.96  percent)  to  give  con- 
siderable support  to  the  average  figure,  9.41  percent.  Serono  and 
Palozzi  (1911)  give  11.05  and  12.09  as  the  minimum  and  maximum 
percents  of  lecithin  in  fresh  yolk.  Tornani  (1909)  says  that  the 
amount  of  lecithin,  and  the  relation  between  cholesterol  and  leci- 
thin in  egg-yolk  is  quite  variable,  that  this  relation  changes  as  the 
eggs  are  kept,  and  is  different  in  fertilized  and  unfertilized  eggs. 
The  amount  found  by  Mesernitzy  (1907)  in  fresh  eggs  was  15.35 
percent  of  the  dry  substance,  and  the  amount  fell  to  about  one-half 
of  that  by  20  days'  incubation.  Glikin  (1908a)  thinks  that  the 
eggs  of  those  species  of  birds  which  are  least  capable  of  independent 
existence  when  hatched  contain  a  larger  supply  of  lecithin  than  oth- 
ers. 

Distinctions  have  been  made  of  several  different  phosphatids  in 
egg-yolk.  Laves  (1903a,  1903b)  states  that  there  are  several  leci- 
thins present,  and  that  they  are  largely  free,  but  partly  in  combina- 
tion with  protein  and  perhaps  partly  with  cerebrin. 


172  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Henriques  and  Hansen  (1903)  found  that  changes  in  the  char- 
acter of  the  fat  of  the  food  did  not  lead  to  much  alteration  in  the  io- 
dine number  of  the  lecithin,  though  it  did  in  that  of  the  egg  fat.  The 
lecithin,  then,  seemed  to  retain  its  constancy  of  composition  inde- 
pendent of  the  food  fat.  Barbieri  (1910,  1912)  made  an  examina- 
tion of  the  extracts  of  egg-yolk  usually  said  to  contain  lecithin,  and 
came  to  the  conclusion  that  the  yolk  contains  no  lecithin,  either  free 
or  combined,  that  is,  that  there  was  no  choline  and  no  phosphorus 
united  to  glycerol.  The  phosphorus  was  said  to  be  dialyzable,  and 
all  or  in  part  in  the  form  of  soluble  phosphates.  We  have  not  seen 
confirmation  of  this  conclusion. 

Stern  and  Thi  erf  elder  (1907)  found  three  distinct  phosphatids 
in  egg-yolk:  the  first,  orange  red,  was  thought  to  be  lecithin;  the 
second,  bright  yellow,  resembled  cephalin ;  and  the  third,  white,  was 
apparently  a  diamino-monophosphatid.  MacLean  (1908b,  1909b, 
1909c)  reports  a  monamino-diphosphatid,  and  Frankel  and  Bolaffio 
(1908),  a  triamino-monophosphatid,  neottin. 

Eppler  (1913)  separated  two  phosphatids  from  egg  yolk.  They 
differ  in  solubility  in  alcohol,  and  in  the  proportion  of  nitrogenous 
base  (choline)  which  is  present. 

OTHER  PHOSPHORUS-CONTAINING  COMPOUNDS  REPORTED  PRESENT  IN  EGGS 

The  substance  to  which  von  Bunge  (1885a)  gave  the  name 
haematogen,  and  which  he  and  Hugounenq  and  Morel  (1905a, 
1905b)  examined  with  especial  view  to  its  being  of  the  nature  of  an 
embryonic  haemoglobin  was  found  to  be  a  pseudonuclein  which  may 
have  been  derived  from  the  vitellin. 

The  mucoid  present  in  egg,  ovomucoid,  has  been  supposed  by 
some  to  contain  phosphorus  (see  Milesi,  1898),  but  if  carefully  pre- 
pared there  is  not  more  than  a  trace  of  phosphorus  (Langstein, 
1903). 

PHOSPHORUS  IN  THE  DIGESTIVE  SECRETIONS 

GASTRIC  JUICE 

In  the  gastric  juice  which  Schoumow-Simanowsky  (1893-94) 
took  by  stomach  fistulae  from  healthy  dogs,  phosphoric  acid  was. 
found  to  the  extent  of  0.00398  and  0.0036  percent. 

Pekelharing  (1902)  believes  pepsin  to  be  free  from  phosphorus, 
and  that  the  phosphorus  which  others  have  found  in  their  prepara- 
tions was  present  as  a  contamination  of  lecithin  or  perhaps  a  phos- 
phorus-containing mucus.  Pekelharing  mentions  Nencki  and  Sieber 
as  believing  that  there  is  a  nucleoprotein  in  the  gastric  juice. 

PANCREATIC  JUICE 

Pancreatic  juice,  caused  to  flow  from  dogs  by  the  introduction 
of  secretin,  is  reported  by  Plimmer  and  Kaya  (1909)  as  showing 
the  following  distribution  of  phosphorus  compounds. 


PHOSPHORUS  METABOLISM 


173 


DISTRIBUTION  OF  PHOSPHORUS  IN  DOGS'  PANCREATIC  JUICE 
Plimmer  and  Kaya  (1909)  Percent  of  Total  Ps05 


Dog 

Ether-sol- 
uble P205 
(lecithin) 

Water-soluble 

P2O5  (nucleic  acid 

and  inorganic) 

Inorganic  P2O5 

Protein  P205 
(nucleoprotein  and 
phosphoprotein  ) 

Phosphopro- 
tein    P2OS 

I 

II 

III 

0 
0 
0 

56.3 
86.3 
59.8 

Present 
Trace 
Trace 

43.7 
13.7 
36.7 

43.7 
13.7 
40.2 

Frouin  and  Gerard  (1912)  determined  the  phosphorus  in  the 
pancreatic  juice  of  the  cow  and  the  dog.  Pancreatic  juice  was  ob- 
tained from  dogs  having  temporary  fistulae  by  the  injection  of  se- 
cretin. From  7  animals  1700  c.c.  of  the  juice  was  collected  in  6-7 
hours.  It  contained  0.006  gm.  P  per  liter.  From  cows  with  per- 
manent fistulae  the  pancreatic  juice  contained  0.0089  gm.  P  per 
liter;  the  juice  as  collected  a  month  later  by  catheter  in  the  pancre- 
atic duct  contained  0.003  gm.  P  per  liter. 

BILE 

The  phosphorus-containing  constituents  of  bile  are  phosphatids 
and  a  very  little  mineral  phosphate.  The  mucus  of  the  liver  secre- 
tion is  said  to  be  either  nucleoalbumin  or  nucleoprotein  (Paijkull, 
1888).  The  inorganic  constituents  of  bile  are  the  chlorides  of  so- 
dium and  potassium  and  small  amounts  of  the  phosphates  of  cal- 
cium, magnesium  and  iron.  Edlefsen  (1880)  taking  the  old  form- 
ula for  haemoglobin,  which  gives  it  a  phosphorus  content,  worked 
out  a  reaction  by  which  the  haemoglobin  of  the  red  corpuscles  dis- 
integrating in  the  liver  may  furnish  a  part  of  the  phosphorus  of 
bile,  a  part  going  to  the  urine. 

The  phosphatids  in  general  are,  of  course,  usually  spoken  of,  in 
all  but  the  more  recent  writings,  as  lecithin,  but  it  is  shown  (Ham- 
marsten)  that  there  are  in  bile  phosphatids  of  different  solubility 
and  different  relation  of  N:P.  Hammarsten  found  evidence  of  a 
jecorin-like  phosphatid,  in  certain  cases  at  least.  The  first  finding 
of  lecithin  in  bile  was  apparently  that  of  Gobley  (1856).  It  was 
then  detected  by  the  presence  of  oleic  acid  and  margaric  acid  in 
some  kind  of  combination  not  a  fat.  Quantitative  determinations 
of  the  lecithin  have  commonly  been  simply  by  computation  from  the 
phosphorus  found  in  the  ether  extract. 

Hammarsten  (1901,  1902,  1904,  1905b)  made  a  study  of  the 
bile  of  different  species  of  animals,  especially  those  of  polar  regions. 
He  notes  that  the  largest  amount  of  lecithin  is  in  the  bile  of  the 
polar  bear  which  uses  a  diet  unusually  rich  in  fat,  and  thinks  it  may 
be  Nature's  provision  for  the  digestion  of  the  fat.  In  the  table 
the  animals  are  arranged  in  order  of  decreasing  phosphatid  content 


174 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


of  the  bile.  The  bile  of  the  liver  and  that  of  the  gall  bladder  are 
said  not  to  be  alike  in  composition,  the  lecithin  being  considerably 
higher  in  the  bladder  bile. 

PHOSPHORUS  FOUND  IN  THE  ETHER  EXTRACT  OF  THE  BILE  OF  VAR- 
IOUS ANIMALS— (Hammarsten,  1905) 


Animal 

Phosphorus  in  ether  extract 
Percent  P 

Lecithin  computed  from  the  same 
Percent 

Dog    

0.911-1.14 
0.048-1.17 
0.100-0.611 

0.768 

0.502 

0.420 

0.334 

0.332 

0.289 

0.272 

0.191 

0.181 

0.168 

0.162 

0.043 

0.033 
Present  but  too  littl 

23.12-28.96 
1.23-29.75 
2.54-15.5 
19.50 

Hog    

12.74 

10.67 

8.47 

8.43 

7.35 

7.04 

Cattle     

4.86 
4.60 

Seal    

4.27 
4.10 

1.08 

Codfish 

0.81 
3  to  be   determined 

Daniel-Brunet  and  Rolland  (1911b)  reported  certain  chemical 
analyses  of  the  bile  and  the  liver  of  cattle.  The  following  table 
shows  the  range  of  the  values  found  for  compounds  of  phosphorus. 

PHOSPHATES,  NUCLEOPROTEINS  AND  LIPOIDS  OF  BILE  OF  CATTLE 

Parts  per  1000 


Phosphates 
P2O5 

Nucleo- 
proteins 

Lipoids 

Total 

Cholesterins 

Lecithins  and 

Min. 

Max. 

Min. 

Max. 

neutral  soaps 

Min. 

Max. 

Min. 

Max. 

Min. 

Max. 

Bile 

1.31 

1.58 

1.15 

2.25 

1.100 

2.130 

0.410 

0.813 

0.690 

1.317 

In  another  article  (1911a)  these  authors  say  that,  with  cattle, 
neither  sex  nor  castration  was  found  to  make  any  difference  as  to 
the  quantity  of  the  mineral  elements,  the  glycogen  or  the  nitrogen- 
ous compounds  in  the  bile;  but  they  did  affect  the  lipoid  content, 
bulls  showing  the  most  lipoids,  cows  less,  and  steers  still  less. 

Rosenbloom  (1912,  1913b)  reports  the  following  analysis  of 
human  bile  taken  from  a  biliary  fistula,  as  parts  per  1000;  total 
solids  29.8,  bile  salts  10.1,  mucin  and  pigments  4.86,  cholesterol  2.61, 
fat  6.85,  soaps  2.6,  lecithin  6.42,  inorganic  matter  9.2,  fatty  acids 
1.2. 

See  also  Brand  (1902),  Yeo  and  Herroun  (1884-5),  Baginsky 
and  Sommerfeld  (1895),  Pruszynski  and  Siemienski  (1906),  Bonan- 
ni  (1906),  and  Long  and  Gephart  (1908b). 


PHOSPHORUS  METABOLISM  175 

SALIVA 

The  saliva  contains  phosphates.  With  regard  to  the  quantita- 
tive relations  of  the  mineral  constituents  of  the  saliva,  the  following 
data  are  at  hand : 

Hammerbacher  (1881)  made  an  organic  and  inorganic  analysis 
of  mixed  human  saliva  in  which  the  inorganic  salts  formed  2.205 
parts  per  1000.      The  ash  analysis  was  as  shown  below: 


Percent 

K20 

45.714 

Na20 

9.593 

CaO  (with  trace  of  iron  oxide) 

5.011 

MgO 

0.155 

so3 

6.380 

PA 

18.848 

CI 

18.352 

104.053 
Oxygen  equivalent  of  chlorine  4.135 


99.918 


By  computation  from  the  ash  analysis,  it  was  concluded  that 
the  ash  of  the  saliva  may  have  been  made  up  of  the  following  com- 
pounds : 

Compound  Percent 

KC1  38.006 

K2S04  13.908 

K3P04  21.278 

Na3P04  16.917 

Ca3(P04)2  9.246 

Mg3(P04)2  0.338 


99.693 
Excess  CI  0.282 


99.975 


This  aggregates  90.109  percent  alkali  salts  and  9.584  percent 
alkaline  earth  phosphates.  Hammerbacher  says  that  Enderlin 
found  92.367  percent  alkali  salts  and  5.509  percent  earth  phosphates 
with  a  trace  of  iron  phosphates. 


0.82 


176  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Schafer's  "Text-Book  of  Physiology,"  Vol.  I,  p.  348  quotes  from 
Jacubowitsch,  through  Hermann's  "Handbuch,"  V  (2),  p.  14,  the 
following  analyses  of  the  ash  from  mixed  saliva,  expressed  as  parts 
per  1000  parts  of  saliva. 

Human  Dog 

Total  solids  1.82  6.79 

Phosphoric  acid  0.51 

Soda  0.43 

Lime  0.03 

Magnesia  0.01  \ 

Alkaline  chlorides  0.84  5.82 

Schafer  also  quotes  (p.  347)  from  Herter,  through  Hoppe-Sey- 
ler,  "Physiol.  Chem.,"  II,  p.  191,  the  following  as  the  salts  found  in 
submaxillary  saliva,  expressed  as  parts  per  1000  of  saliva: 

K2S04  0.209 

KC1  0.940 

NaCl  1.546 

Na2C03  0.902 

CaC03  0.150 

Ca3(POJ2  0.113 

F.  N.  Schulz  gives,  in  Oppenheimer's  "Handbuch  der  Biochemie 

des  Menschen  und  der  Tiere,"  III,  p.  29,  the  following  ash  analyses 

from  Jacobi  (Diss.,  Wurzburg,  1896)  : 

Percent  Percent 

I  II 

CI  14.46  13.68 

K  35.88  35.69 

Na  32.91  21.61 

PA  10.98  17.70 

Sulphuric  acid  Trace        '  7.1 

Ca  2.19  3.96 

Mg  0.47  0.69 

Barille  (1911)  considers  that  dental  tartar  is  derived  from  the 
saliva  through  the  precipitation  of  its  salt  as  tricalcic  phosphate 
and  calcium  carbonate  as  a  result  of  the  loss  of  carbon  dioxide  from 
the  carbonophosphate  in  the  saliva.  Barille  found  in  the  inorganic 
part  of  tartar  70  percent  of  the  phosphate  and  8  percent  of  the  car- 
bonate of  calcium. 

Roger  (1908)  believes  that  the  presence  of  phosphates  is  neces- 
sary for  the  amylolytic  action  of  saliva.  He  showed  that  the  sugar- 
forming  power  of  saliva  may  be  destroyed  by  precipitation  of  the 
phosphates  with  uranium  acetate,  and  restored  by  addition  of  so- 
dium phosphate. 


PHOSPHORUS  METABOLISM  177 

PHOSPHORUS  PRESENT  IN  CHYLE 

Hamill  (1906-07)  collected  chyle  through  a  fistula  in  the  thigh 
of  a  human  subject,  and  examined  it  for  various  constituents.  The 
lecithin  averaged  4.204  gm.  per  100  gm.  of  ether  extract.  Lipase 
and  amylase  were  both  present.  Lecithin  given  by  mouth  during 
the  time  of  observation  produced  a  rise  in  the  ether-soluble  phos- 
phorus of  the  chyle. 

PHOSPHORUS  PRESENT  IN  LYMPH 

From  the  few  phosphorus  estimations  on  lymph  which  have  come 
to  our  attention  we  are  unable  to  conclude  as  to  its  usual  content. 
Odenius  and  Lang  (1874)  found  13  parts  per  1000  of  soluble  P205; 
E.  Ludwig  (quoted  by  von  Zeynek,  1895)  found  15  parts  total  P203 
per  1000;  von  Zeynek  (1895)  found  0.095  parts  soluble,  and  0.214 
parts  insoluble  P205  per  1000  c.  c,  while  Zaribnicky  (1912)  found 
in  the  lymph  of  the  horse  0.089  parts  total  P205  per  1000.  The 
preceding  figures  were  from  analyses  of  human  lymph.  See  also 
Dahnhardt  (1866). 


178  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


PART  IV 

NORMAL  PHOSPHORUS  METABOLISM 
MAINTENANCE  OF  NEUTRALITY 

The  approximate  constancy  of  reaction  of  the  body  fluids, 
which  is  one  of  the  conditions  essential  to  life,  is  maintained  at  neu- 
trality (actually  very  slight  alkalinity)  by  a  group  of  counteracting 
agencies  prominent  among  which  are  the  phosphates. 

The  need  for  such  an  adjustment  is  due  to  the  facts  that  the 
predominating  chemical  reaction  of  the  body  is  the  oxidation  of  car- 
bon compounds  to  carbonic  acid,  and  that  sulphuric  and  phosphoric 
acids  are  prominent  among  the  normal  products  of  protein  katabo- 
lism. 

This  control  of  reaction  in  the  body  fluids  is  accomplished  by 
phosphates,  carbonic  acid  and  carbonates,  aided  by  the  amphoteric 
proteins,  and  sustained  by  the  excretory  functions  of  the  lungs  for 
carbonic  acid,  and  of  the  kidneys  and  intestine  for  phosphates. 

We  shall  consider  in  detail  only  Henderson's  theory  of  the  pe- 
culiar effectiveness  of  carbonic  and  phosphoric  acids  and  their  salts 
in  this  adjustment.  (Henderson,  L.  J.,  1906,  1098a,  1908b,  1908c, 
1909,  1911,  1913 ;  Henderson  and  Black,  1907,  1908 ;  Fitz,  Alsberg 
and  Henderson,  1907.)  Henderson's  explanation  of  this  phenom- 
enon is  based  on  physico-chemical  considerations  and  ion  concen- 
tration measurements.  The  substances  considered  are  the  two 
pairs  of  compounds,  carbonic  acid  and  bicarbonate,  and  the  mono- 
and  di-phosphate.  The  processes  concerned  may  be  represented  by 
the  two  equations, 

M,HP04+HA^MA+MH2P04 
MHC03+HA?±MA+H2C03 

where  M  stands  for  any  basic  radical ;  A,  for  any  acid  radical.  The 
indications  are  that  in  the  fluids  of  the  body  the  carbonic  acid  is 
present  as  carbonic  acid  (H2C03)  or  sodium  bicarbonate  (NaHC03), 
and  all  of  the  phosphoric  acid  as  mono-sodium  phosphate 
(NaH2P04)  or  di-sodium  phosphate  (Na2HP04).  Now,  mixtures  of 
these  compounds  in  solution  possess  greater  power  than  any  other 
known  salts  for  balancing  each  other,  and  any  acid  or  base  that  may 
be  added,  so  that  the  solution  remains  neutral.  While  a  pure  solution 


PHOSPHORUS  METABOLISM  179 

of  NaH2P04  is  very  weakly  acid,  and  one  of  Na2HP04  is  very  weak- 
ly alkaline,  one  of  a  mixture  of  the  two  adjusts  itself  by  ionization 
so  that  it  is  neutral ;  and  addition  of  an  acid  to  such  a  solution  does 
not  give  it  even  the  slight  acidity  of  mono-sodium  phosphate  till 
enough  acid  has  been  added  to  convert  all  of  the  di-sodium  salt  to 
mono-sodium  salt.  Bases  produce  an  opposite  effect,  but  will  not 
give  the  mixture  even  the  faint  alkaline  reaction  of  di-sodium  phos- 
phate till  all  has  been  converted  to  that  form. 

The  case  is  similar  for  carbonic  acid  and  bicarbonates,  and 
when  all  four  compounds  are  present  in  a  solution,  the  base  dis- 
tributes among  these  components  so  that  the  relative  amounts  of 
each  are  constant  for  any  given  temperature. 

By  mathematical  deductions  Henderson  shows  that  the 
strength  of  an  acid  theoretically  best  fitted  to  preserve  neutrality 
in  a  pure  solution  is  such  that  its  ionization  constant  is  near  IX 10"7, 
which  is  the  ionization  constant  of  distilled  water;  or,  more  exactly, 
"that  its  ionization  constant,  divided  by  the  degree  of  ionization  of 
its  salt,  is  precisely  equal  to  the  hydrogen  ion  concentration  in  pure 
water."  The  ionization  constant  of  H2C03  is  3X10"T>  and  that  of 
the  H2P04~  ion  is  2X10"7*  Hence  both  of  these  are  theoretically, 
as  they  are  found  to  be  practically,  nearly  ideal  neutralizing  agents 
near  the  neutral  point. 

The  condition  to  be  maintained,  however,  is  not  exactly  the  neu- 
trality of  distilled  water,  and  at  the  alkalinity  required  very  slight 
variations  in  hydrogen  ion  concentration  are  accompanied  by  great- 
er variations  in  sodium  bicarbonate  content  than  in  the  ratio  of 
mono-  and  di-sodium  phosphate.  Hence  the  carbonate  is  more  effi- 
cient than  the  phosphate,  proportionately,  in  maintaining  alkalinity. 
The  concentration  of  bicarbonate  is  then  from  ten  to  twenty  times 
the  concentration  of  carbonic  acid,  and  neutrality  is  attained  only 
when  the  concentration  of  bicarbonate  has  fallen  nearly  to  that  of 
the  carbonic  acid.  This  efficiency  of  the  bicarbonate-carbonic  acid 
equilibrium  is  further  increased  by  the  maintenance  of  a  nearly  con- 
stant supply  of  carbonic  acid  through  its  removal  by  the  lungs. 

The  phosphates,  however,  have  certain  other  advantages.  At 
the  most  critical  point,  that  is,  when  the  bicarbonate  is  nearly  used 
up  and  the  system  is  very  near  the  neutral  point,  the  phosphates  are 
most  effective.  Furthermore,  they  are  present,  especially  in  proto- 
plasm, probably  in  much  greater  amounts  than  the  carbonate,  and 
are  correspondingly  of  greater  service.  In  addition,  the  easy  dif- 
f  usibility  of  acid  phosphates  and  their  removal  by  the  excretory  or- 
gans must  greatly  enhance  their  practical  efficiency. 


180  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

With  regard  to  the  magnitude  of  the  effect  of  these  two  agents 
together  in  the  protoplasm,  Henderson  says:  "Only  when  an  amount 
of  acid  equal  to  about  three-fourths  of  the  total  carbonic  acid  plus 
the  total  phosphoric  acid  of  the  protoplasm  has  been  added  to  the 
protoplasm  can  there  be  a  real  beginning  of  acidity." 

This  mechanism  of  equilibrium  in  the  tissues  and  fluids  very 
greatly  reduces  the  alkali  requirement  of  the  body;  but  it  must  be 
sustained,  as  it  is,  in  fact,  by  special  retention  mechanisms  at  the 
points  of  excretion,  by  means  of  which  the  alkalis  are  returned  to 
the  blood  after  the  excretion  by  lungs  and  kidneys  of  acids  with 
which  these  bases  were  combined. 

In  so  far  as  the  alkalis  are  united  with  carbonic  acid  they  are 
entirely  saved  to  the  body,  as  the  carbonic  acid  leaves  the  body  in 
the  gaseous  state  through  respiration.  The  amount  of  alkali  in  use 
in  this  way  is  said  to  be  about  1.5  gm.  at  any  moment,  or  400  gm. 
per  day. 

Alkalis  are,  however,  removed  from  the  body  in  the  urine  of 
omnivora  and  carnivora,  chiefly  as  phosphates  and  sulphates.  At 
the  kidney  there  are  two  provisions,  probably  about  equal  in  their 
effect,  for  sparing  alkali.  First,  the  fixed  alkalis  are  replaced  by 
ammonia,  in  so  far  as  it  is  available.  This  is  all  clear  gain  of  base, 
because  the  ammonia  comes  from  nitrogenous  compounds  which  are 
nearly  neutral,  and  if  completely  oxidized  becomes  neutral  urea.  The 
other  factor  which  saves  alkalis  to  the  body  is  the  physico-chemical 
process  by  which  acid  urine  is  separated  from  the  alkaline  blood. 
Though  the  change  of  reaction  is  small,  the  amount  of  base  involved 
may  be  considerable  in  the  presence  of  phosphates  and  other  salts 
and  acids  having  the  ability  to  undergo  wide  variation  in  the 
amount  of  base  which  they  hold  with  but  slight  variation  in  hydro- 
gen ion  concentration.  As  we  have  already  noted,  phosphates  are 
peculiarly  effective  in  this  way  near  the  neutral  point.  Carbonic 
acid  and  uric  acid  have  the  same  property,  and,  at  hydrogen  ion  con- 
centrations such  as  are  reached  in  acidosis,  the  /?-oxybutyric  acid 
and  its  salts,  which  are  then  present,  are  also  effective  in  the  same 
way. 

With  regard  to  the  relative  magnitude  of  the  parts  played  by 
the  proteins,  the  bicarbonates  and  the  phosphates  in  maintaining 
neutrality  nothing  definite  can  be  asserted,  nor  can  it  be  said  that 
there  are  not  other  unrecognized  agencies  involved.  Conditions 
within  the  body  cannot  be  exactly  duplicated  outside,  especially  as 
to  the  influence  of  colloids;  but  as  far  as  true  solutions  are  con- 
cerned quantitative  measurements  can  be  made  with  pure  substan- 


PHOSPHOEUS  METABOLISM  181 

ces  which  give  some  idea  of  the  possible  degree  of  effectiveness  of 
the  different  agents;  and  the  final  word  in  the  discussion  between 
T.  Brailsford  Robertson  and  Lawrence  J.  Henderson  (Robertson, 
1909b,  1910a;  Henderson,  L.  J.,  1908a,  1908c,  1909)  seems  to  be 
that,  so  far  as  the  blood  is  concerned,  in  passing  from  the  reaction 
of  normal  blood  to  that  of  blood  in  advanced  acid  intoxication,  the 
proteins  of  the  blood  are  about  one-fifth  as  efficient  as  the  bicar- 
bonates  in  maintaining  its  neutrality.  In  the  tissues  or  tissue- 
fluids  the  proteins  very  likely  play  a  larger  part.  Henderson  attrib- 
utes a  relatively  greater  effectiveness  to  bicarbonate  than  to  phos- 
phate except  at  points  very  near  to  neutrality. 

Michaelis  (1913)  gives  the  average  H+  ion  concentration  of  hu- 
man venous  blood,  in  normal  resting  condition,  as  2.75  X10"8  at 
20-21°  and  4.46X10"8  at  37.5° ;  and  the  greatest  variations  from  this 
mean,  found  under  such  conditions,  amounted  to  -{-12  and  — 11.7 
percent  of  the  mean.  A  difference  of  ±  6  percent  is  attributable  to 
uncertainties  of  the  method  of  determination,  so  that  the  variations 
observed  are  but  very  slightly  beyond  the  limits  of  error  of  work. 

THE  ABSORPTION  AND  ELIMINATION  OF  COMPOUNDS  OF 

PHOSPHORUS 

GENERAL  CONSIDERATIONS 

After  the  ingestion  of  ordinary  food,  with  its  variety  of  phos- 
phorus compounds,  organic  and  inorganic,  there  begins  at  once  the 
same  process  of  simplification  and  absorption  that  is  general  for 
other  groups  of  its  nutrient  components.  The  inorganic  phosphates 
are,  of  course,  absorbed  as  such.  The  organic  phosphorus  com- 
pounds are  absorbed  in  part  without  change,  in  part  after  partial 
cleavage,  and  in  part  after  complete  separation  into  their  simplest 
groups,  with  the  liberation  of  phosphorus  as  inorganic  phosphoric 
acid,  the  method  differing  with  the  nature  of  the  nutrient.  The 
phosphoric  acid  split  off  from  organic  compounds  during  digestion 
behaves  just  as  does  any  other  inorganic  phosphoric  acid  introduced 
as  such  (Oeri,  1909).  The  larger  part,  though  by  no  means  all  of 
the  food  phosphorus,  therefore,  is  absorbed  as  inorganic  phosphate. 
For  details  as  to  the  digestion  of  the  several  groups  of  phosphorus 
compounds  see  the  special  discussions  of  these  groups. 

Such  inorganic  phosphates  as  are  insoluble  in  water  are  most 
of  them  readily  soluble  in  the  hydrochloric  acid  of  the  gastric  juice. 
They  are  then  absorbed,  mainly  in  the  small  intestine,  which  is  usu- 
ally faintly  acid  in  the  upper  portions  (Raudnitz,  1893 ;  and  Wuertz, 
1912). 


182  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

It  is  usually  said  to  be  true  that  absorption  of  phosphates  takes 
place  only  in  the  acid  portion  of  the  small  intestine,  but  von  Wendt 
and  Zuckmayer,  as  noted  below,  suggest  means  by  which  phos- 
phates may  be  absorbed  from  the  alkaline  portions  of  the  ali- 
mentary tract. 

Since  the  phosphorus  of  the  food  is  mostly  in  completely  oxi- 
dized forms  there  can  be  no  significant  change  in  the  state  of  oxi- 
dation during  metabolism.  There  is  evidence  of  some  capacity,  how- 
ever, for  animals  to  oxidize  the  less  completely  oxidized  compounds, 
as  is  observed  by  Heffter  (1903),  who  states  that  in  the  healthy  or- 
ganism phosphorous  acid  is  completely  oxidized.  He  states,  also, 
that  pyrophosphates  and  hypophosphites  are  excreted  unchanged, 
while  metaphosphates  are  changed  to  the  ortho-  form. 

The  circumstances  which  determine  whether  phosphates  shall 
be  eliminated  through  the  urine  or  the  feces  are  first,  the  reaction 
of  the  contents  of  the  alimentary  tract,  as  determined  especially  by 
hydrochloric  acid  and  carbonates;  second,  the  nature  and  relative 
amounts  of  the  other  mineral  elements,  especially  the  bases,  present 
in  the  digestive  tract,  as  affecting  the  solubility  of  the  phosphates ; 
third,  after  the  absorption  of  the  phosphates  from  the  alimentary 
tract,  all  of  those  circumstances  which  control  the  secretion  of  phos- 
phates into  the  digestive  tract  and  the  reabsorption  from  the  same ; 
fourth,  the  many  circumstances  which  affect  the  reaction  of  the 
blood  serum  and  the  amounts  and  proportions  of  the  other  salts 
present,  as  affecting  the  form  and  therefore  the  solubility  Of  the 
phosphates,  the  more  readily  soluble  tending  to  go  out  through  the 
urine;  and  fifth,  the  species  of  animal  involved. 

There  may  be  a  considerable  variation  as  to  path  of  outgo 
without  apparent  change  of  conditions;  thus  the  experiments  of 
Kochmann  (1911)  (see  Inn.  of  Amount  of  Food  on  P.  Met.)  show 
that  on  the  same  amounts  of  the  same  kind  of  food  there  may  be  a 
change  of  urine  phosphorus  from  27  to  34.9  percent  of  the  total 
outgo,  and  of  feces  phosphorus  from  65.1  to  73  percent  of  the  total 
outgo,  without  apparent  cause. 

This  whole  problem,  then,  is  almost  hopelessly  complicated, 
though  we  are  able  to  record  a  considerable  number  of  harmonious 
observations  as  to  the  bearing  of  certain  individual  factors. 

The  general  course  of  the  minerals  during  their  passage  through 
the  alimentary  tract  has  been  traced  by  Wildt  (1874,  1879),  who 
made  a  study  of  digestion  in  sheep  by  analyses  of  the  contents  of 
the  different  parts  of  the  alimentary  tract.  By  means  of  quanti- 
tative estimations  of  the  silica  and  also  the  absorbable  constituents 
of  the  food,  in  each  part  of  the  tract,  it  was  possible  to  trace  absorp- 
tion, to  recognize  both  the  organic  and  inorganic  constituents  of 


PHOSPHORUS  METABOLISM 


183 


each  of  the  digestive  secretions,  and  later  to  tell  where,  when,  and  to 
what  extent  these  constituents  were  again  absorbed  and  returned  to 
the  blood  stream. 

The  amount  of  phosphorus  secreted  into  the  alimentary  tract 
was,  in  each  of  the  three  sheep  studied,  greater  than  the  total 
amount  in  the  food.  Secretion  of  phosphorus  greatly  exceeded  ab- 
sorption in  the  first  and  second  stomachs,  but  was  less  than  absorp- 
tion in  the  third  stomach ;  changed  again  sO  that  secretion  exceeded 
absorption  in  the  abomasum  and  duodenum,  and  once  more  so  that 
absorption  exceeded  secretion  in  the  remainder  of  the  tract. 

The  principal  secretion  of  phosphorus  was  in  the  saliva  and  in 
the  secretion  of  the  abomasum,  and  its  largest  absorption  was  from 
the  small  intestine.  Calcium  and  magnesium,  on  the  other  hand, 
were  secreted  principally  into  the  small  intestine,  and  were  absorbed 
most  actively  from  the  first  and  second  stomachs  and  the  colon. 

The  following  tables  set  forth  the  facts  as  Wildt  determined 
them  as  to  the  absorption  and  secretion  of  phosphorus  and  the  min- 
eral bases. 

ABSORPTION  AND   SECRETION  OF   PHOSPHORUS,   CHLORINE   AND 

THE  MINERAL  BASES  IN  THE  ALIMENTARY  TRACT  OF 

THREE  SHEEP— Grams 


- 

First  and 

second 
stomachs 

Third 
stomach 

Abomasum 

and 
duodenum 

Small 
intestine 

Caecum 

Colon 

Large  intestine 

Entire 

Last  part 

Sheep      I 

K           "II 

III 

—2.525 
+1.993 
—2.104 

-2.232 
-4.811 
-2.480 

+1.665 
+5.218 
+9.683 

+3.746 
-4.714 
-6.955 

—7.436 
—3.321 
—2.073 

-0.391 
—1.607 
-0.903 

-0.186 

0.525 

-1.042 

+2.178 
+0.761 
-0.912 

Sheep      I 

Na           '       II 

III 

+14.794 
+15.673 
+13.006 

-10.004 
-10.167 
-6.389 

+2.983 

+7.167 

+20.715 

+15.871 
+7.060 
-1.687 

—14.837 
-12.566 
—17.728 

-3.527 
-1.746 
-2.967 

-3.560 
-3.086 
-3.740 

-3.179 
-1.703 
-1.935 

Sheep      I 

Ca          "        II 

III 

-0.729 
—0.236 
-1.002 

+0.163 
+0.214 

+0.278 

-1.284 
-0.803 
+0.536 

+1.415 
+0.118 
-0.328 

+0.653 
+0.398 
+0.754 

-1.023 
-0.686 
-0.585 

+0.555 
+0.107 
+0.050 

-0.424 
-0.386 
+0.198 

Sheep      I 

Mg"       "        II 

"      III 

-0.675 
-0.472 
—0.724 

-0.316 
-0.128 
-0.189 

+0.109 
—0.129 
+0.596 

+0.719 
+0.557 
-0.199 

-0.175 
—0.235 
+0.328 

-0.479 
-0.274 
-0.278 

+0.022 
-0.289 
+0.089 

+0.207 
+0.311 
-0.010 

Sheep      I 

P2O5      ' '        II 

III 

+7.359 
+9.064 
+5.416 

-2.068 
—2.861 
-1.091 

+0.155 
+2.818 
+8.939 

+3.470 
-2.164 
-8.701 

-7.348 
—5.175 
—2.440 

-0.523 
-0.642 
-0.437 

-0.707 
-0.290 
-0.416 

+0.980 
-0.353 
-0.556 

Sheep       I 

CI        "11 

"      III 

—1.284 
+1.821 
—1.538 

+0.925 
—1.425 
—1.012 

+12.835 

+18? 

+27.422 

—1.687 
—10.619? 
-16.032 

—9.558 
-7.167 
-9.584 

-1.209 
-0.150 
-1.389 

-1.491 
-1.141 

-0.828 
-2.174 

( — )==more  absorbed  than  secreted.       (  +  )=more  secreted  than  absorbed. 

SECRETION  DURING  TWENTY-FOUR  HOURS  IN  VARIOUS  PARTS  OF 
THE  ALIMENTARY  CANAL— Grams 


Mouth   (Saliva) 

Abomasum 

Small  Intestine 

Total 

DailyFood 

K    

Mg 
PsOB'.... 

15.010 
7.969 

4.443 
7.291 

17.240 
2.584 

9.270 
0.601 

4.443 

31.571 

0.601 

17.240 

10.553 

9.078 
1.103 
3.267 

3.162 
0.966 

184  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

One  point  which  a  study  of  these  tables  makes  clear  is  the 
utter  absurdity  of  the  usual  statements  as  to  digestibility  of  min- 
eral nutrients,  and  also  of  many  statements  as  to  coefficients  of  ab- 
sorption of  inorganic  salts. 

The  mineral  salts,  including  phosphates,  reach  the  general  cir- 
culation almost  wholly  through  the  portal  system.  The  final  dis- 
position of  the  phosphates  under  ordinary  conditions  of  nutrition 
may  then  involve  retention  in  the  body,  or  elimination,  either  in 
urine  or  feces.  There  are  also  the  various  losses  of  phosphorus  in- 
volved in  reproduction  and  lactation.  It  would  seem  from  the  work 
of  Taylor  (1911)  that  there  is  no  cutaneous  elimination  of  phos- 
phates. Taylor  studied  the  cutaneous  elimination  of  nitrogen,  sul- 
phur and  phosphorus  with  two  laboratory  assistants  in  periods  of 
28  and  45  days.  The  average  daily  cutaneous  eliminations  were  as 
follows : 

Substance  Subject  J.  Subject   D. 

Grams  Grama 

S  0.028  0.015 

P  0.003  0.002 

N  0.190  ,  0.160 

The  author  concluded  that  the  phosphorus  figure  stands  for 
desquamation  only,  and  that  there  is  not  a  real  cutaneous  elimina- 
tion of  phosphorus  in  any  form. 

In  discussing  urinary  and  feces  excretion  of  calcium  salts, 
VonWendt  (1905)  suggests  that  the  acidity  of  the  urine  is  respon- 
sible for  its  content  of  calcium  salts,  while  the  calcium  salts  which 
are  excreted  into  the  intestine  are  rendered  more  or  less  insoluble  by 
the  alkalinity  of  the  intestinal  juices,  their  reabsorption  being  there- 
by restricted.  Phosphates  are  supposed  to  be  excreted  into  the  in- 
testine as  mono-  and  di-salts.  Here  the  di-salt  decomposes  into 
mono-  and  tri-  salts,  and  the  latter,  further,  with  the  taking  up  of 
the  elements  of  water,  into  basic  phosphate  and  acid.  Since,  of 
these  phosphates,  onlythe  mono-salt  can  be  considered  as  relatively 
easily  soluble  in  the  weakly  alkaline  intestinal  juice,  their  reabsorp- 
tion takes  place  principally  in  this  form,  though  to  some  extent  also 
as  the  di-salt.  From  the  relation  of  Ca:P  in  the  feces  vonWendt 
concludes  that  the  calcium  is  present  as  the  trimetallic  phosphate. 

Zuckmayer  (1912)  found  that  "tricalcol,"  an  alkali-soluble  col- 
loidal calcium  preparation,  was  absorbed  from  the  small  intestine 
of  a  rabbit  and  a  dog,  while  the  tricalcium  phosphate,  directly  intro- 
duced, was  not  usually  absorbed.  Zuckmayer  states  that  in  nor- 
mal digestion  calcium  (calcium  phosphate)  is  withdrawn  from  food 


PHOSPHORUS  METABOLISM  185 

substances  by  the  acid  of  the  stomach,  with  the  formation  of  acid 
calcium  phosphate  and  calcium  chloride,  and  probably  also  phos- 
phoric acid  in  the  presence  of  protein  cleavage  products.  He  also 
suggests  that  in  the  small  intestine,  as  soon  as  there  is  sufficient  al- 
kali present,  the  dissolved  calcium  phosphates  are  again  changed  to 
tricalcic  phosphate,  which  becomes  absorbable  through  the  agency 
of  the  colloidal  protein  cleavage  products. 

In  studying  the  conditions  governing  calcium  and  phosphorus 
metabolism  in  infants  Bliihdorn  (1912-13)  experimented  with  cal- 
cium and  phosphorus  compounds  added  to  feces  extracts  in  vitro. 
If  to  8  c.c.  of  distilled  water  were  added  1  c.c.  each  of  N/50 
Na2HP04  and  N/100  Ca  Cl2  a  slight  but  distinct  precipitate  of 
Ca,  (P04)  2  appeared.  By  similar  addition  of  these  salt  solutions  to 
water-extracts  of  feces  it  was  determined  that  if  Ca  salts  occur  with 
phosphates  in  the  intestine,  in  weakly  acid  or  alkaline  reaction,  in- 
soluble Ca3(P04)2  results,  and  in  this  way  Ca  and  P  may  be  with- 
drawn from  absorption.  A  strong  acid  reaction  prevented  the  pre- 
cipitation of  Ca3(P04)2.  The  colloids  of  the  feces  extracts  appeared 
to  play  no  decisive  role  in  these  processes. 

Plimmer  (1913a)  studied  the  action,  on  phosphorus  compounds, 
of  enzymes  obtained  from  pancreas,  liver,  intestine,  castor  beans, 
yeast  and  wheat  bran.  The  phosphorized  nutrients  studied  includ- 
ed glycerophosphoric  acid,  phytic  acid,  nucleic  acid,  casein  and 
others.  The  most  active  tissue  in  the  hydrolysis  of  the  organic 
phosphorus  compounds  was  thought  to  be  the  intestinal  mucous 
membrane.  Phytic  acid  was  attacked  readily  only  by  an  enzyme  in 
the  bran  extract.  Casein  was  the  only  one  of  these  compounds 
which  was  hydrolyzed  by  the  pancreas  extract.  From  the  activity 
of  the  intestinal  mucosa  in  the  cleavage  of  organic  phosphorus  com- 
pounds Plimmer  concludes  that  they  are  absorbed  only  in  the  hy- 
drolyzed condition,  the  phosphorus  as  inorganic  phosphate.  The 
lipoclastic  enzymes  were  shown  not  to  be  active  in  the  hydrolysis 
of  organic  phosphorus  compounds. 

While  Plimmer's  conclusion  as  to  the  absorption  of  phosphorus 
compounds  only  in  a  state  of  hydrolytic  cleavage  is  doubtless  true 
of  the  bulk  of  the  phosphorus  absorbed  from  organic  compounds,  his 
evidence  is  not  of  such  nature  as  to  controvert  successfully  the  es- 
tablished facts  as  to  the  absorption  of  a  part  of  the  organic  phos- 
phorus of  the  food  in  unsplit  or  incompletely  hydrolyzed  forms. 


186  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

THE  INFLUENCE  OF  PHOSPHATES  ON  DIGESTION 

The  nature  of  the  influence  of  phosphates  in  digestion  was 
shown  by  Berg  and  Gies  (1906-7),  who  studied  the  effects  of  ions 
on  catalysis,  with  especial  reference  to  peptolysis  and  tryptolysis. 
The  H+  ion  is  the  favorable  factor  in  peptolysis,  and  the  OH"  ion  in 
tryptolysis ;  the  associated  ions,  or  molecules,  or  both,  exerting 
deterrent  influences  in  variable  degrees.  Variations  in  swelling  ef- 
fects on  fibrin,  in  general,  correspond  to  these  digestive  differences. 

Finzi  (1903)  determined  that  intravenous  injection  of  neutral 
sodium  phosphate  caused  increased  secretion  of  saliva,  and  in- 
creased phosphate  content  of  the  saliva. 

Ferrata  and  Moruzzi  (1907)  determined  that  a  phosphorus-con- 
taining food  causes  an  increase  in  the  lecithin  content  of  the  intes- 
tinal mucous  membrane  of  the  dog,  while  the  nucleoproteids  were 
increased  after  the  taking  of  food  of  any  kind. 

Roger  (1908)  found  that  the  amylolytic  power  of  the  saliva  de- 
creased with  the  addition  of  uranium  acetate,  and  disappeared  with 
the  complete  precipitation  of  the  salivary  phosphates.  The  ad- 
dition of  sodium  phosphate  then  caused  the  amylolytic  action  to  re- 
appear.     Conclusions  were  sustained  by  adequate  numerical  data. 

Giacosa  and  Dezani  (1909)  found  phosphates  in  the  press-juice 
from  pig  stomach,  but  also  prepared  from  it  a  phosphorus-free  di- 
gestive enzyme. 

A.  Loeb  (1910)  showed  with  a  dog  that  the  urinary  phospho- 
rus was  reduced,  temporarily,  by  the  taking  of  food,  apparently  be- 
cause of  its  utilization  in  the  preparation  of  the  digestive  juices. 

Lisbonne  (1911)  reports  that  phosphates  do  not  activate  salt- 
free  amylases  of  the  saliva  and  pancreatic  secretion  as  do  chlorides. 

W.  Lob  (1911)  demonstrated  that  phosphates  stand  in  an  im- 
portant relation  to  the  utilization  of  sugar.  Glycolysis,  which  is 
favored  by  hydroxyl  ions,  is  accelerated  by  the  presence  of  phos- 
phate ions;  with  constant  hydroxyl  ion  concentration,  the  acceler- 
ation rising  with  the  absolute  amount  of  phosphate  present.  Leci- 
thin or  glycerophosphates  hinder  phosphate  glycolysis. 

The  activating  effect  of  primary  and  secondary  phosphates  on 
various  proteases  is  compared  by  Fernbach  and  Schoen  (1911). 

The  above  notes  show  that  phosphates  stand  in  such  a  signifi- 
cant relation  to  the  digestion  and  utilization  of  food  that  their  ex- 
cretion into  the  alimentary  tract  must  be  regarded  as  of  vital  con- 
sequence. 

The  possible  significance  of  phosphates  in  relation  to  enzymatic 
processes  in  animal  bodies  is  indicated  by  a  series  of  studies  by 
Wroblewski,  Harden,  Young,  and  Euler  and  associates,  on  the  rela- 
tion of  phosphates  to  the  enzymes  of  yeast. 


PHOSPHORUS  METABOLISM  187 

Wroblewski.(1901)  demonstrated  the  favorable  effect  of  a  certain 
concentration  of  phosphate  on  the  action  of  zymase  from  yeast 
press-juice.  Harden  and  Young  (1906,  1908a,  1908b,  1909,  1910, 
1911a,  1911b)  also  found  that  phosphate  accelerates  the  fermenta- 
tion of  sugar  by  yeast  juice,  the  phosphate  becoming  non-precip- 
itable  by  magnesia  mixture.  Young  (1907,  1909, 1911)  shows  that 
the  organic  phosphorus  compound  above  mentioned  is  a  hexose  phos- 
phate. The  method  of  action  of  the  enzyme  involved,  phosphatese, 
is  discussed  in  a  series  of  papers  by  Euler  and  Kullberg  (1911), 
Euler  and  Ohlsen  (1911),  Euler  and  Lundeqvist  (1911),  Euler  and 
Ohlsen  (1912),  Euler  (1912a)  and  Euler  and  Backstrom  (1912). 
Other  studies  on  the  influence  of  phosphates  on  the  enzymes  of 
yeast  are  those  of  Euler  and  Beth  af  Ugglas  (1911). 

AUTOLYSIS  OF  COMPOUNDS  OF  PHOSPHORUS 

The  intermediary  metabolism  of  phosphorus  is  accomplished 
through  the  agency  of  enzymes.  Given  the  particular  con- 
ditions necessary  for  their  activity,  they  are  able  to  bring  about  the 
various  changes  in  the  state  of  organization  of  matter  which  are  es- 
sential to  the  maintenance  of  life.  Our  state  of  knowledge  of  these 
compounds  is  especially  incomplete,  and  we  make  no  effort  to  pre- 
sent a  full  discussion  of  the  subject.  We  merely  offer  a  few  notes 
on  investigations  demonstrating  the  existence  of  such  compounds, 
and  the  nature  of  the  cleavage  which  they  may  induce;  and  we 
make  the  suggestion  that  reversibility  of  direction  is  a  not  infre- 
quent characteristic  of  enzyme  action,  some  at  least  of  the  autolytic 
enzymes  undoubtedly  possessing,  with  change  of  conditions,  capac- 
ity for  synthesis  in  the  same  field  of  activity.  Evidence  of  the  ac- 
tivity of  autolytic  enzymes  has  been  accumulated  by  a  great  number 
of  workers,  among  whom  are  those  whose  names  follow,  these  hav- 
ing given  attention  to  compounds  of  phosphorus. 

Kutscher  and  Lohmann  (1903)  found  no  autolytic  decomposi- 
tion of  the  lecithin  of  the  brain  of  the  ox,  but  Coriat  (1904a)  found 
an  enzyme  in  brain  tissue  capable  of  splitting  choline  from  lecithin. 
This  enzyme  acts  only  in  neutral  or  slightly  alkaline  media,  and  the 
yield  of  choline  is  greater  in  the  latter  than  in  the  former. 

Satta  (1907,  1908)  determined  that  both  lecithin  and  nuclein 
phosphorus  in  ox  liver,  pancreas  and  thymus  split  off  inorganic 
phosphorus  in  autolysis,  the  nuclein  more  rapidly  than  the  lecithin. 
Below  are  data  from  this  study : 


188  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

THE  AUTOLYSIS  OF  ORGANIC  PHOSPHORUS  COMPOUNDS 

Satta  (1907,  1908) 


Animal 

Total  phos- 
phorus as  P2O5 

Percent 

Percents  of  total  phosphorus 

Organ 

A  lcohol-ether-sol- 

uble  (Lecithin, 

jecorin,  etc.) 

Water-  and  acetic 
acid-soluble 
(Inorganic) 

Freed  by 

ashing: 
(Nuclein) 

of 

autolysis 
Days 

Ox 
Ox 
Ox 
Ox 
Ox 

2.9 

2.8 
2.7 
2.65 
2.89 

30.60 

28.2 

25.20 

23.60 

21.61 

31.79 

51.8 

62.95 

66.46 

69.9 

37.61 
20.0 
11.85 
9.94 
8.49 

2 

7 

12 

21 

Ox 
Ox 
Ox 

3.87 
3.23 
3.47 

41.69 
34.20 
11.47 

22.71 
60.50 
84.73 

35.60 
5.30 
3.80 

i 
5 

Calf 
Calf 
Calf 
Calf 
Calf 

3.9 
4.0 
3.8 
3.8 
3.8 

10.95 
9.56 
9.26 
9.1 
9.0 

20.05 
24.27 
43.72 
51.75 
59.46 

69.00 
66.17 
,47.02 
39.15 
31.54 

1 

3 

10 

30 

Lymph,  sarcoma 
of  intestine.... 

Man 

1.85 

19.66 

39.59 

40.75 

Satta  and  Fasiani  (1910)  determined  that  the  presence  of  lip- 
oids increased  the  autolysis  of  liver,  as  measured  by  the  amount  of 
nitrogen  passing  into  solution  in  a  given  time.  The  action  seemed 
to  be  most  pronounced  when  the  liver  of  a  starved  animal  was  used 
for  the  autolysis.  The  action  was  not  proportional  to  the  amount 
of  lipoids  present. 

Simon  (1911)  demonstrated  active  autolysis  in  brain  tissue, 
which  affected  not  only  the  phosphorus  compounds  soluble  in  alco- 
hol and  ether,  but  also  those  which  are  insoluble,  the  proportion  of 
the  former  to  the  latter,  represented  by  the  inorganic  phosphorus 
split  off  from  organic  compounds,  being  as  61  to  39. 

Grund  (1912a,  1912b,  1913)  determined  that  in  degenerating 
muscle,  following  the  division  of  its  nerves,  there  occurs  a  large  in- 
crease of  fat,  and  of  water  in  proportion  to  fat-free  substance ;  but 
in  the  dry,  fat-free  substance  the  total  nitrogen,  total  phosphorus 
and  phosphatid  phosphorus  remain  constant.  On  the  other  hand, 
the  ratio  of  protein  phosphorus  to  total  phosphorus  greatly  increas- 
es, as  also  the  ratio  of  protein  phosphorus  to  protein  nitrogen.  Thus 
the  protein  phosphorus  maintains  its  integrity,  while  phosphorus- 
free  proteins  dissolve.  Degeneration  as  caused  by  hunger  is  ac- 
companied by  the  katabolism  of  all  the  protein  components. 

For  a  discussion  of  autolysis  of  the  several  groups  of  organic 
phosphorus  compounds  see  the  discussions  of  the  metabolism  of  the 
same. 

METHOD  OF  URINARY  ELIMINATION  OF  PHOSPHATES 

In  view  of  the  importance  of  the  function  of  the  kidneys  in 
freeing  the  system  from  certain  classes  of  katabolites,  and  thus 
assisting  in  the  maintenance  of  that  constancy  of  conditions  in  the 


PHOSPHORUS  METABOLISM  189 

blood  which  is.  essential  to  the  vital  functions,  it  is  a  matter  of  in- 
terest to  know  the  method  of  formation  of  urine  and  the  agencies 
through  which  this  is  accomplished. 

M.  Maly  (1876)  shows  that  acid  constituents  diffuse  and  di- 
alyze  more  rapidly  than  alkali  from  such  mixtures  as  are  found  in 
the  blood,  this  depending  on  the  greater  mobility  of  ionized  hydro- 
gen, and  of  more  complex  substances  which  contain  hydrogen,  than 
of  corresponding  basic  substances.  Thus  acid  phosphates  are  sep- 
arated from  alkaline  blood. 

This  theory  receives  support  from  the  investigation  of  Fitz, 
Alsberg  and  Henderson  (1907),  who  demonstrated  by  hydrochloric 
acid  feeding  to  rabbits  that  the  resultant  shifting  of  the  salt  equi- 
librium toward  the  acid  side  increased  the  urinary  elimination  of 
phosphates. 

In  harmony  also  with  this  idea  is  the  observation  of  Teissier 
(1877)  of  inverse  relations  in  the  excretion  of  glucose  and  phos- 
phates, the  significance  of  which  probably  lies  in  an  accentuation  of 
the  state  of  acidosis  during  diminished  glucose  outgo. 

Liebermann  (1893b)  found  that  alkaline  solutions  passed 
through  a  filter  of  lecithalbumin  become  acid.  He  therefore  ad- 
vanced the  theory  (with  experiments)  that  acid  urine  may  arise  in 
such  a  way,  by  the  acid  bodies  in  the  kidney  cells  removing  base 
from  alkaline  urates  as  the  urine  passes,  provided  there  is  not  too 
much  alkali. 

Frey  (1911)  also  investigated  the  nature  of  the  process  by 
which  phosphorus  is  separated  from  the  blood  in  the  kidneys.  By 
intravenous  injection  of  Na2HP04,  and  phosphorus  determinations 
on  the  blood  serum  and  urine,  he  found  that  the  urine  contained 
from  10  to  60  or  70  times  the  concentration  of  phosphorus  that  ex- 
ists in  blood  serum ;  thus  it  appears  that  the  process  of  separation 
of  phosphorus  from  the  blood  in  the  kidneys  is  more  than  one  of 
filtration. 

Folin  (1905)  in  discussing  the  laws  governing  the  compo- 
sition of  the  urine  maintains  the  incorrectness  of  the  usual  view 
(see,  for  instance,  G.  Zuelzer,  1905  and  Ott,  1886)  that  the  phos- 
phates of  normal  acid  human  urine  are  present  to  the  extent  of 
about  60  percent  in  the  diacid  form,  and  states  that  the  phosphates 
of  clear  acid  urine  are  all  monobasic,  the  acidity  of  such  urines  be- 
ing ordinarily  greater  than  the  acidity  of  all  the  phosphates,  the  ex- 
cess being  due  to  free  organic  acids. 

It  is  Folin's  idea  that  the  precipitate  with  barium  chloride 
which  is  ordinarily  considered  to  include  the  diacid-phosphates  in 
reality  contains  phosphorus  only  as  an  impurity  in  the  abundant 
precipitate  of  barium  sulphate. 


190  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

According  to  Hammarsten  (1911),  p.  784,  phosphorus  may  be 
found  in  urinary  sediments  as  tricalcic  and  trimagnesium  phos- 
phates in  alkaline  urines,  as  dicalcium  phosphate  in  neutral  or  faint- 
ly acid  urine,  and  as  ammonium  magnesium  phosphate  in  urines 
which  have  become  ammoniacal  through  alkaline  fermentation,  or 
in  amphoteric  urines  in  the  presence  of  a  sufficient  quantity  of  am- 
monium salts. 

ORGANIC  PHOSPHORUS  IN  THE  URINE 

From  an  early  date  it  has  been  recognized  that  a  portion  of  the 
phosphorus  of  the  urine  is  present  in  organic  combination.  Ronalds 
(1846)  considered  that  he  found  organic  phosphorus  in  the  urine, 
though  he  states  that  his  methods  were  unsatisfactory. 

Sotnitschewsky  (1880)  determined  glycerophosphoric  acid  as  a 
common  constituent  of  human  urine,  though  the  amount  present  is 
sometimes  very  small. 

W.  Zuelzer  (1881a)  found  that  the  urine  of  a  man  22-25  years 
old  contained  1-2  mg.  of  phosphorus  (stated  as  P205)  combined  with 
glycerin.  In  fever  no  more  was  found,  but  much  more  was  found 
after  chloroform  anaesthesia,  subcutaneous  injection  of  morphine, 
after  the  crisis  of  pneumonia,  and  in  erysipelas.  The  maximum 
amount  found  by  Zuelzer  for  24  hours  is  35  mg.  P205. 

Lepine  and  Eymonnet  (1882)  examined  the  urine  of  more  than 
100  subjects.  Normal  human  urine  they  found  to  contain  about. 
0.150  gm.  of  glycerophosphoric  acid  per  liter,  being  about  0.15  to 
0.3  of  one  percent  of  the  amount  of  the  nitrogen.  With  fatty 
degeneration  of  the  liver  in  tuberculosis  the  value  of  the  glycero- 
phosphoric acid,  in  percent  of  nitrogen,  was  found  to  be  1.0-1.8,  or 
about  6  or  7  times  the  normal. 

In  a  later  article  (Lepine,  Eymonnet  and  Aubert,  1884)  they  re- 
port increased  proportions  of  unoxidized  to  total  phosphoric  acid  in 
the  urine  in  apoplexy,  epilepsy  and  delirium  tremens,  and  in  a  dog 
after  subcutaneous  injection  of  hydrochlorate  of  morphine,  and  also 
in  a  dog  after  the  ingestion  of  potassium  bromide.  They  state  that 
in  pernicious  anaemia  it  may  be  present  in  four  times  the  normal 
amount,  without  an  increase  in  the  total  phosphorus.  An  excess 
above  normal  occurs  also  in  some  cases  of  icterus,  typhoid  fever  and 
acute  pneumonia ;  while  in  cases  of  meningitis  the  unoxidized  phos- 
phorus was  present  in  less  than  the  normal  relation  to  the  nitro- 
gen of  the  urine. 

Biilow  (1894)  found,  with  dogs,  that  the  phosphoric  acid  esters 
of  the  urine  did  not  vary  greatly  with  the  introduction,  either  per  os 
or  subcutaneously,  of  glycerophosphates,  these  salts,   in  so  far  as 


PHOSPHOEUS  METABOLISM  191 

they  were  excreted  in  the  urine,  being  mostly  decomposed  to  phos- 
phates.     His  figures  are  as  follows : 

URINARY  EXCRETION  OF  PHOSPHORIC  ACID  ESTERS  BY  DOGS 


Days 

H3P04  as  ether-phosphoric  acid 

5 

0.00619 

Normal    conditions 

1 

0.01085 

3  gm.  calcium  glycerophosphate-)- 2  gin, 
calcium  carbonate  per  os 

1 

0.01191 

3  gm.  sodium  glycerophosphate,  suhcutane- 
ously 

1 

0.00626 

Next   day 

1 

0.00573 

3   gm.   salol 

5 

0.00626 

Next  day 

Rockwood  (1895)  showed,  by  the  method  of  Siegfried,  that 
normal  urine  contains  both  carnic  and  phosphocarnic  acids.  He 
found,  as  did  Siegfried,  two  compounds  in  the  iron  precipitate,  one 
"carniferrin,"  the  iron  compound  of  phosphocarnic  acid,  soluble  in 
alkalis,  and  the  other  a  basic  iron  compound  of  carnic  acid. 

Ceconi  (1896)  states  that  the  organic  phosphorus  of  the  urine 
equals  11-28  mg.  per  day,  the  amount  varying  with  the  volume;  that 
values  greater  than  20  mg.  are  found  only  in  pathological  condi- 
tions ;  and  that  the  organic  phosphorus  of  the  urine  is  not  affected 
by  variations  in  the  food. 

Oertel  (1898-9)  found,  in  a  study  of  the  urinary  phosphorus  of 
7  healthy  men,  an  average  total  phosphorus  content  of  2.0  gm.  P205, 
and  about  0.05  gm.  P205  as  organic  phosphorus.  The  highest 
amount  of  organic  phosphorus  for  one  day  was  0.120,  and  the  low- 
est 0.03  gm.  P205.  The  amount  was  not  influenced  by  work,  but 
differed  much  with  individuals. 

Keller  (1900a)  determined  that  the  organic  phosphorus  of  the 
urine  of  infants  was  greater  in  amount  after  feeding  cow's  milk 
than  after  feeding  human  milk,  though  it  was  a  smaller  percentage 
of  the  total  urinary  phosphorus,  since  the  total  phosphorus  from 
the  cow's  milk  greatly  exceeded  that  resulting  from  woman's  milk. 
It  varied  in  amount  from  less  than  one  to  about  10  percent  of  the  to- 
tal urinary  phosphorus. 

In  a  fasting  experiment  on  himself  Keller  found  in  a  four-day 
test  that  the  amount  increased  daily,  thus,  0.017,  0.029,  0.034,  and 
0.057  gm.  P,05.  These  increased  amounts  constituted  increased 
percentages  of  the  total  urinary  phosphorus,  though  this  also  in- 
creased from  day  to  day. 

The  significance  of  organic  phosphorus  of  the  urine  was  not  es- 
tablished. It  did  not  vary  regularly  with  the  phosphorus  given  in 
the  food,  nor  with  absorbed  phosphorus,  nor  with  nitrogen  metabol- 
ized. It  was  not  influenced  by  Na,HP04  ingested.  On  a  similar 
diet  normal  children  excreted  more  organic  phosphorus  than  did 


192  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

children  suffering  from  such  disturbance  as  would  reduce  powers  of 
oxidation.  The  amount  eliminated  was  not  altered  by  feeding  with 
a  very  low  phosphorus  diet. 

Lepine  (1901)  states  that  in  fatty  degeneration  of  the  liver  its 
high  lecithin  content  is  accompanied  by  an  abnormal  glycerophos- 
phate acid  content  of  the  urine,  which  may  reach  a  value  as  high 
as  twice  the  usual  amount.  He  suggests  that  an  abundance  of  in- 
completely oxidized  phosphorus  in  urine  may  be  an  indication  of  a 
fatty  condition  of  the  liver. 

Mandel  and  Oertel  (1902)  observed  no  change  in  the  organic 
phosphorus  of  the  urine  by  food  rich  in  organic  phosphorus. 

Y.  Henderson  and  Edwards  (1903)  compared  phosphorus  fig- 
ures on  the  urine  by  direct  titration  and  after  fusion.  From  the 
correspondence  of  results  they  conclude  that  no  organic  phosphorus 
was  present. 

K.  Bornstein  (1905)  considered  the  origin  of  the  organic  phos- 
phorus of  the  urine,  in  metabolism  experiments  on  himself.  Over- 
feeding with  protein  did  not  increase  the  organic  phosphorus  of  the 
urine.  The  author  did  not  determine  the  source  of  this  organic 
phosphorus  but  was  of  the  opinion  that  it  was  of  endogenous  origin. 

Symmers  (1904-5a,  1904-5b)  has  investigated  the  elimination 
of  organic  phosphorus  in  the  urine  in  various  pathological  condi- 
tions. Symmers  calls  attention  to  the  idea,  which  has  prevailed  un- 
til recently,  that  the  urinary  excretion  of  organic  phosphorus,  eith- 
er in  health  or  in  disease,  is  negligible,  and  cites  8  earlier  investi- 
gations of  the  subject.  Symmers  states  that  exercise  and  diet  do 
not  affect  the  urinary  excretion  of  organic  phosphorus.  Analyses 
of  the  urine  of  20  cases,  embracing  9  diseases,  are  given.  The  or- 
ganic phosphorus  varies  from  a  small  part  to  about  nine-tenths  of 
the  total  phosphorus.  Symmers  cautions,  therefore,  against  the 
use  of  inorganic  phosphorus  determinations  as  indices  of  metab- 
olism. He  finds  the  excretion  of  organic  phosphorus  to  some  ex- 
tent rhythmical,periods  of  excessive  secretion  alternating  with  what 
may  be  either  retention  or  diminished  production.  Symmers  finds 
organic  phosphorus  elimination  pronounced  in  lymphatic  leukaemia ; 
and  also  in  nervous  diseases  of  degenerative  type,  to  such  extent  as 
could  not  be  derived  from  the  destruction  of  nervous  tissue. 

In  conclusion  Symmers  says  that  abnormal  increase  of  organic 
phosphorus  may  be  explained  either  as  an  increase  in  the  production 
of  phosphorized  endogenous  metabolic  products,  or  as  an  expression 
of  lessened  oxidation,  with  the  organic  phosphorus  compounds  as 
the  end-products. 


PHOSPHORUS  METABOLISM  193 

Symmers  determined  total  phosphorus  by  titration  with  urani- 
um nitrate,  after  fusion;  and  inorganic  phosphate  also  by  uranium 
nitrate  titration,  but  without  fusion.  Organic  phosphorus  was 
reckoned  by  difference. 

Franchini  (1907,  1908a)  found  glycerophosphoric  acid  quite 
variable  in  amount  in  the  urine  of  fasting  rabbits.  When  lecithin 
was  fed,the  average  glycerophosphoric  acid  content  of  the  urine  was 
increased,  but  there  was  no  uniform  effect  noticeable.  No  choline 
or  formic  acid  was  found  as  a  decomposition  product  from  lecithin. 

Plimmer,  Dick  and  Lieb  (1909-10)  found  the  inorganic  phospho- 
rus of  the  urine  to  constitute  90-100  percent  of  the  whole,  its 
amount  depending  especially  on  the  intake  of  P205.  Organic  phos- 
phorus excretion  they  found  very  irregular  and  not  dependent  on 
the  diet.  They  concluded,  therefore,  that  it  must  be  of  endogenous 
origin. 

Mathison  (1910)  determined  organic  phosphorus  in  the  urine  of 
5  healthy  persons  on  3-6  consecutive  days,  and  in  the  urine  of  one 
person  as  affected  by  exercise  and  by  the  ingestion  of  glycerophos- 
phoric acid  and  sodium  glycerophosphate.  Organic  phosphorus  was 
found  normally  present,  usually  in  amounts  greater  than  0.1  gm. 
P205  per  day,  though  occasionally  it  fell  below  this  figure,  and  in  one 
case  reached  0.3  gm.  The  proportion  of  the  total  phosphorus  pres- 
ent in  organic  combination  varied  considerably  from  day  to  day  but 
averaged  about  6  percent. 

The  organic  phosphorus  outgo  was  not  affected  by  ingestion  of 
glycerophosphoric  acid.  The  organic  phosphorus  of  urine  was 
found  readily  dialysable  and  not  precipitable  by  reagents  which  pre- 
cipitate protein. 

Kondo  (1910)  studied  the  effect  of  organic  phosphorus  of  the 
food  on  the  organic  phosphorus  of  the  urine.  With  a  dog  weighing 
8  kg.  the  urinary  organic  phosphorus  varied  from  0.0086  to  0.0227 
gm.  P205  per  day,  while  the  total  urinary  phosphorus  varied  from 
0.3941-1.6436  gm.  P205  daily.  The  variations  in  organic  phospho- 
rus, then,  were  slight  in  absolute  quantity.  The  proportion  of  or- 
ganic to  inorganic  phosphorus  was  less  (1.2-1.6  percent)  on  days  of 
excessive  phosphorus  ingestion,  in  the  shape  of  brain,  casein  and 
thymus,  than  on  intermediate  days  (2.2-3.5  percent),  when  the 
phosphorus  was  present  in  horse  flesh. 

Bogdanow  (1911)  found  the  organic  phosphorus  of  the  urine 
to  rise  significantly  immediately  after  the  fall  in  temperature  in 
croupous  pneumonia. 


194 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  6 


To  summarize  the  evidence:  Urine  may  contain  organic  phos- 
phorus as  glycerophosphoric  acid,  and  as  phosphocarnic  acid.  This 
fraction  is  variable  in  amount  from  a  very  small  to  a  large  part  of 
the  whole,  and  is  too  large  a  factor  to  ignore  in  any  quantitative 
work.  This  organic  phosphorus  increases  after  fast  (Keller), 
chloroform  anaesthesia,  morphine  injection,  and  in  many  pathologi- 
cal conditions.  Its  significance  is  not  known.  Its  amount  seems 
slightly  to  increase  after  the  ingestion  of  glycerophosphates,  but 
there  is  certainly  no  marked  or  constant  effect  of  the  constituents 
of  the  food,  either  phosphorus-containing  or  otherwise,  or  of  the 
amount  of  phosphorus  absorbed,  or  of  the  nitrogen  metabolism,  on 
this  excretion.  It  appears  to  be  to  some  extent  an  individual  char- 
acteristic, though  temporary  physiological  states  seem  to  be  the 
dominant  factors  in  its  quantitative  variation. 

PHOSPHORUS  COMPOUNDS  IN  THE  FECES 

The  phosphorus  compounds  of  the  feces  are  of  mixed  origin, 
being  derived  in  part  each  from  food  residues,  intestinal  excretions, 
digestive  secretions,  intestinal  epithelium,  and  bacterial  products, 
the  relative  proportion  of  the  whole  which  is  contributed  by  each 
of  these  factors  varying  much  with  the  nature  of  the  diet  and  kind 
of  animal,  the  feces  being  the  main  vehicle  of  outgo  for  metabolized 
as  well  as  unmetabolized  phosphorus  in  herbivora,  while  among 
carnivora  and  omnivora  the  main  outgo  of  metabolized  phosphorus 
is  through  the  urine.  Since  there  is  no  evidence  of  a  direct  rela- 
tionship between  food  and  feces  phosphorus  compounds,  compara- 
tively little  attention  has  been  given  the  determination  of  the  ident- 
ity of  the  compounds  in  the  feces.  Inorganic  salts  occur  in  abund- 
ance both  as  intestinal  secretions  and  as  food  residues  after  more  or 
less  reapportionment  of  bases  during  the  digestive  process. 

The  following  analyses  of  the  ash  of  meconium  are  quoted  from 
F.  Miiller  (1884).  Analyses  I,  II,  III  and  IV  are  of  human  mecon- 
ium and  were  quoted  by  Miiller  from  Zweifel. 

ASH  ANALYSES  OF  MECONIUM— Percent  of  Ash 


I 

II 

III 

Horse 

Human 

Human 

Human 

Human 

0.30 

0.67 

0.80 

0.87 

1.36 

2.60 

0.86 

18.76 

8.00 

31.80 

5.70 

5.09 

2.65 

4.32 

3.60 

4.00 

7.23 

10.21 

10.66 

7.80 

5.40 

3.20 

38.42 

47.05 

22.30 

23.00 

39.50 

21.92 

24.42 

K  6.00 
Na  24.20 

... 

8.40 

3.78 

2.53 

8.68 

IV 
Human 


Insol.    in   HC1 

Fe203    

CaO    

MgO     

P2Os 

SOa     

Alkalis    

CI     


0.80 

9.50 

7.92 

8.58 

81.90 

Ns  15.93 

K7.09 

8.90 


PHOSPHORUS  METABOLISM  195 

Feces  from  long  fasting  were  shown  to  resemble  meconium. 
Flesh  feces  also  resemble  meconium,  and  seem  to  consist  largely  of 
excretion  products  of  the  intestinal  canal  rather  than  of  food  resi- 
dues. 

Muller  also  states  that  the  time  of  passage  of  the  contents  of 
the  alimentary  tract  affects  its  composition ;  thus,  if  the  movement 
be  rapid  one  finds  unaltered  bile  salts  and  abundant  alkali  salts,  but 
if  the  feces  remain  in  the  large  intestine  for  a  week  or  more  almost 
no  alkali  compounds  appear  in  the  feces  as  passed. 

Micko  (1900),  studying  feces  phosphorus  with  adult  human  be- 
ings, concludes  that  the  organic  phosphorus  of  the  feces  does  not 
come  for  the  most  part  from  the  food,  but  that  some  true  nuclein 
is  present  in  the  feces  from  an  ordinary  mixed  diet  and  also  in  the 
feces  from  diets  of  flesh  and  plasmon. 

In  a  later  paper  (1900)  Micko,  Muller,  Poda  and  Prausnitz  con- 
clude that  human  feces  are  derived  in  large  part,  with  but  few  ex- 
ceptions, from  intestinal  secretions,  and  (Muller)  that  cow's  milk 
used  by  either  infants  or  adults  leaves  no  phosphorus-rich  casein 
residue  in  the  feces. 

Schilling  (1901a)  states  that  when  the  food  contains  only  a 
small  amount  of  phosphorus  the  feces  contain  little  or  no  crystalline 
tricalcium  phosphate. 

Von  Oefele  (1901)  states  that  in  diabetes  and  syncopic  neurosis 
the  relative  scarcity  of  triple  phosphate  crystals  in  the  feces  can 
be  explained  as  due  to  coexistent  phosphaturia.  The  presence  of 
triple  phosphates  in  the  feces  was  found  to  have  no  especial  diagnos- 
tic value. 

A  number  of  workers  have  determined  lecithin  in  the  feces. 
Among  these,  Deucher  (1898)  determined  lecithin  in  the  feces  in 
cases  of  occlusion  of  the  pancreatic  duct;  P.  Muller  (1900)  found  lec- 
ithin, calculated  as  distearyl-lecithirt,  in  milk  feces,  to  the  extent  of 
1.13-10.1  percent  of  the  ether  extract,  and  considered  it  as  derived 
largely  from  the  food. 

Long  (1906b)  and  Johnston  (L.  &  J.,  1906,  1907)  have  studied 
the  phosphorus  of  the  feces  fat.  Ether-soluble  phosphorus  com- 
pounds which  they  speak  of  as  lecithins,  are  found  in  amounts  of 
from  one  to  several  percent,  but  they  suggest  that  these  may  be  but 
remotely  related  to  simple  distearyl  lecithin.  They  consider  that 
this  fraction  may  come  in  part  each  from  food,  intestinal 
epithelium,  bacteria  and  bile  residues.  Commenting  on  the  usual 
belief  that  lecithin  can  not  escape  digestion,  since  it  is  easily  split 
by  the  pancreatic  secretion,  they  suggest  that,  while  this  may  be 
true  for  the  largest  part  of  the  food  lecithins,  it  may  not  be  equally 
true  of  the  lecithins  of  the  bile  under  certain  conditions. 


196 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


Franchini  (1907, 1908a)  determined  lecithin  in  the  feces  of  rab- 
bits. There  was  no  uniform  increase  produced  by  the  feeding  of 
lecithin  during  fast,  but  the  average  lecithin  content  of  the  feces 
was  greater. 

Peritz  (1908-9b)  reports  lecithin  in  the  feces  in  cases  of  tabes 
and  paralysis  as  varying  between  0.1311  and  7.221  gm.  per  day, 
there  being  no  evidence  of  relationship  between  these  amounts  and 
the  content  of  the  food  in  lecithin. 

A.  Bornstein  (1909)  found  0.02  gm.  lecithin  per  day  in  the 
feces  from  a  diet  of  crackers  and  milk. 

In  connection  with  a  phosphorus  metabolism  study  Rogozinski 
(1910)  made  separations  of  the  groups  of  phosphorus  compounds 
in  the  feces  of  a  man  and  of  two  dogs.  The  rations  and  feces  phos- 
phorus data  were  as  indicated  below. 

PHOSPHORUS  COMPOUNDS  IN  HUMAN  AND  IN  DOG  FECES 
Percent  of  Total  P  O 


Period 

(5 
days) 

Day 

Diet 


Lecithin 


Phytin 


Inorganic 


Protein 


SERIES  I  DOG 


Meat,  rice,  pork  fat    

Meat,  rice,  pork  fat,  phytin 

Meat,  rice,  pork  fat    

Meat,  rice,  lecithin    

Meat,   rice,  pork  fat 

Meat,  rice,  pork  fat,   Na  phosphate 
Meat,  rice,  pork  fat    

SERIES  II  DOG 

Meat,   rice,  pork  fat  

Meat,  rice,  pork  fat,  phytin    

Meat,   rice,  pork  fat  

Meat,   rice,  lecithin    

Meat,   rice,  pork   fat  

Meat,  rice,  pork  fat,  Na  phosphate 

Meat,  rice,  pork  fat 

SERIES  ni  MAN 

Mixed  diet    

Mixed  diet    

Mixed   diet    

Mixed  diet    

Mixed   diet    . 

Mixed  diet,  phytin 

Mixed  diet,  phytin 

Mixed  diet,  phytin 

Mixed  diet,  phytin 

Mixed  diet,  phytin 

Mixed   diet    

Mixed   diet    

Mixed   diet    

Mixed  diet    

Mixed  diet    


1.82 

6.34 

51.00 

0.45 

69.62 

14.25 

1.46 

4.04 

47.52 

2.19 

6.18 

48.69 

1.20 

10.27 

39.74 

1.16 

8.92 

50.43 

1.29 

6.84 

40.15 

40.84 
15.68 
46.98 
42.94 
48.79 
39.49 
51.72 


1.68 

26.16 

42.76 

0.36 

62.42 

14.71 

1.71 

18.72 

40.68 

5.06 

13.53 

40.21 

2.11 

20.47 

39.94 

1.40 

22.84 

43.54 

1.92 

21.61 

43.13 

29.40 
22.51 
38.89 
41.20 
37.48 
32.22 
33.34 


6 

7 
8 
9 

10 


11 
12 
13 
14 
15 


20.46 

3.15 

43.82 

20.18 

4.17 

46.59 

18.19 

3.01 

50.51 

21.88 

0.00 

44.87 

19.49 

1.58 

44.80 

17.24 

2.66 

14.96 

2.45 

16.95 

1.49 

24.05 

0.00 

19.52 

0.00 

16.90 
20.45 
21.56 
14.80 
20.47 


0.00 
3.43 
2.24 
0.00 
0.44 


45.67 
50.77 
52.88 
52.87 
54.38 


53.97 
50.69 
49.93 
55.47 
47.38 


32.57 
29.06 
28.29 
33.25 
34.13 


34.43 
31.82 
28.68 
23.08 
26.10 


29.13 
25.43 
26.27 
29.73 
81.71 


PHOSPHORUS  METABOLISM  197 

The  group  of  compounds  designated  "phytin"  is  that  fraction 
soluble  in  acidified  water;  protein  phosphorus  is  considered  to  be 
that  portion  of  the  organic  phosphorus  which  is  insoluble  in  acidi- 
fied water. 

In  dog  feces  inorganic  and  protein  phosphorus  predominate, 
though  there  is  also  considerable  phytin  phosphorus,  especially 
when  phytin  is  fed.  The  feeding  of  lecithin  also  increased  the  feces 
lecithin. 

In  human  feces  there  is  a  very  much  larger  proportion  of  leci- 
thin phosphorus  than  in  dog  feces.  The  phytin  is  much  more  com- 
pletely absorbed  by  the  human  being  than  by  the  dog,  and  the  feces 
phytin  is  not  increased  in  human  beings  by  the  ingestion  of  phytin. 
This  difference  in  the  utilization  of  phytin  is  in  harmony  with  the 
natural  differences  of  the  diets  of  the  two  species.  The  dog  has  no 
need  for  capacity  to  digest  phytin,  since  there  is  no  phytin  in  its 
natural  food. 

Stutzer  (1908)  found,  in  feeding  experiments  with  sheep  on 
hay,  that  the  feces  contained  metabolic  organic  phosphorus  com- 
pounds insoluble  in  acid  gastric  juice  in  quantities  greater  than 
would  have  been  expected  from  the  analysis  of  the  food. 

Emmett  (1909)  determined  phosphorus  on  the  ether  extract  of 
pig  feces,  and  found  0.39-0.74  percent  P205  in  this  extract. 

Lipschiitz  (1910b)  determined  that  the  phosphorus  content  of 
the  feces  of  young  dogs  on  mother's  milk,  cow's  milk,  meat  diet, 
and  phosphorus-poor  diet  varied  between  0.87  percent  with  mother's 
milk  and  0.60  with  meat  diet,  while  the  starvation  feces  contained 
about  the  same  (0.63  percent)  phosphorus  content.  The  phospho- 
rus intake  with  cow's  milk  was  0.201  gm.  per  kg.  live  weight ;  with 
phosphorus-poor  diet  0.023-0.031  gm.,  and  on  meat  diet  0.095  gm., 
from  which  it  would  appear  that  the  phosphorus  content  of  dog 
feces  under  these  conditions  originated,  not  in  the  food,  but  in  the 
secretions  of  the  digestive  tract. 

Lipschiitz  quotes  figures  for  the  phosphorus  content  of  feces 
as  follows :  Blauberg— infants  6-7  days  old,  0.26-0.49  percent ;  Tiger- 
stedt — man  on  low-phosphorus  diet,  0.9  percent ;  Miiller — man,  fast- 
ing, 1.8-2.2  percent,  and  dog,  grown,  fasting,  1.91  percent;  and 
Munk — dog,  grown,  fasting,  3.65  percent,  from  which  he  concludes 
that  the  feces  arising  from  the  digestive  tract  of  the  growing  organ- 
ism is  notably  poorer  in  phosphorus  than  that  of  adults. 

Fraps  and  Rather  (1912)  have  investigated  the  composition 
and  digestibility  of  the  ether  extract  of  a  variety  of  hays  and  fod- 
ders, and  of  certain  constituents,  among  others  phosphoric  acid,  of 


198  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

the  ether  extract.  Since  the  ether  extract  of  the  feces  is  to  a  con- 
siderable extent  a  bile  residue,  and  not  directly  related  to  the  food, 
it  seems  to  us  that  their  statements  as  to  the  relative  digestibility 
of  the  lecithin  and  other  constituents  of  the  ether  extract  are  quite 
without  warrant. 

VonWendt  (1905)  and  Zuckmayer  (1912)  both  concluded  that 
phosphorus  is  in  the  feces  largely  in  the  form  of  the  trimetallic 
phosphates.     See  p.  184. 

Summary.  The  phosphorus  compounds  of  the  feces  are  not 
well-known,  but  have  been  found  to  include  nuclein  phosphorus,  in- 
organic phosphates  in  a  variety  of  forms,  lecithin-like  compounds 
and  other  organic  metabolic  products,  the  nature  of  which  has  not 
been  investigated.  Studies  of  the  feces  in  fasting,  and  during  feed- 
ing on  meat  and  milk,  lead  to  the  belief  that  a  considerable  part  of 
the  feces  phosphorus,  especially  the  lecithin-like  compounds,  have 
an  origin  other  than  in  the  food.  To  relate  the  feces  phosphorus  com- 
pounds as  a  whole  directly  to  those  of  the  food,  as  in  computation  of 
digestibility,  is  without  justification. 

PHOSPHORUS  EXCRETION  AS  AFFECTED  BY  ACIDS,  SALTS, 
GENERAL  CHARACTER  OF  DIET,  AND  SPECIES  OF  SUBJECT 

ACIDS 

One  of  the  most  potent  influences  affecting  the  elimination  of 
phosphates  is  the  ingestion  of  acids  and  acid  salts.  Under  this 
influence  calcium  and  phosphorus,  being  associated  in  the  body,  are 
affected  at  the  same  time,  and  thus  are  excreted  in  combination;  in- 
creasing acidity  of  the  urine  being  associated  with  increased  excre- 
tion of  both  calcium  and  phosphorus. 

Schetelig  (1880)  studied  urinary  calcium  under  different  condi- 
tions with  a  normal  subject  and  24  cases  of  a  number  of  diseases. 
Incidentally  phosphorus  was  considered.  The  ingestion  of  hydro- 
chloric acid  caused  an  increase  of  both  calcium  and  phosphorus  in 
the  urine. 

Ott  (1886)  reported  the  acid  and  neutral  phosphates  of  human 
urine  as  being  in  the  relation  60:40.  He  determined  that  cal- 
cium is  present  in  urine  in  greater  amount  than  the  solubility  of  its 
acid  and  neutral  phosphates  in  water  would  explain,  and  that  the 
solubility  of  CaHP04  is  increased  by  the  presence  of  acid  phosphates 
of  the  alkalis,  gypsum,  ammonium  chloride  and  magnesium  sul- 
phate. Neutral  alkali  phosphates  decrease  the  solubility  of  tricalcio 
phosphate. 

Rudel  (1893a),  in  experiments  on  dogs  and  infants,  proved  that 
ingestion  of  hydrochloric  acid,  calcium  acetate,  or  chalk,  would  in- 
crease the  urinary  calcium,  while  sodium  phosphate  was  without 


PHOSPHORUS  METABOLISM 


199 


such  effect,  and  'calcium  phosphate  served  markedly  to  reduce  the 
urinary  calcium.  Subcutaneous  injection  of  calcium  acetate  caused 
increased  urinary  calcium  as  also  did  prolonged  peristalsis  from 
tinct.  opii. 

Rudel  (1893b)  showed  with  a  nine-months-old  infant  that  the 
introduction  of  sodium  phosphate  into  the  food  would  decrease  the 
calcium  to  41.76-61.5  percent  of  the  normal.  A  notable  increase  in 
the  urinary  calcium  followed  the  administration  of  dilute  hydro- 
chloric acid. 

In  harmony  with  these  observations  is  that  of  Gerhardt  and 
Schlesinger  (1899)  that  the  administration  of  sodium  bicarbonate 
markedly  reduced  the  urinary  calcium  and  correspondingly  in- 
creased the  feces  calcium.  Similarly  Proskauer  (1910)  found  an 
evident  increase  of  calcium  and  a  smaller  increase  of  magnesium 
in  infant's  blood  in  severe  digestive  disturbance.  In  harmony  with 
this  is  the  observation  of  Allers  and  Bondi  (1907)  of  an  increase  of 
calcium  in  the  blood  in  experimental  hydrochloric  acid  poisoning, 
from  0.069  gm.  to  0.159  gm.  in  1000  gm.  of  blood. 

W.  Camerer,  Jr.  (1902b),  from  a  study  of  conditions  affecting 
ammonia  excretion,  submits  data  on  the  relative  amounts  of  total 
phosphorus,  and  phosphorus  in  acid  salts,  in  the  urine,  as  affected 
by  age,  sex,  exercise,  nature  of  the  food,  and  the  ingestion  of  acid 
and  alkaline  compounds.      The  figures  are  as  follows: 

AVERAGE  DAILY  TOTAL  PHOSPHORUS,  AND  PHOSPHORUS  IN  ACID 
SALTS,  IN  URINE— Grams 


^^m  ■ 

Grown 

men  on 

mixed 

diet 

Youths 

Children 

Men 

Sp.  gr. 

1.016 

Women 
Sp.  gr. 

1.011 

Animal 
food 

Mixed 
diet  plus 
4-5  gm. 

HCl 

Mixed 

Resting 

Exer- 
cise 

Sp.  gr. 
1.0165 

Sp.  gr. 
1.0225 

diet  plus 

8  gm. 
NaHCO 

Total  P2O5 

P2O5  in  acid 

2.89 
1.70 

1.80 
1.01 

1.80 
0.82 

0.70 
0.38 

1.08 
0.64 

2.07 
1.31 

1.56 
1.14 

3.5 
1.55 

1.79 
1.08 

2.635 
0  70 

Here  we  note  a  much  higher  proportion  of  acid  salts  resulting 
from  the  ingestion  of  HCl  than  from  NaHC03. 

Folin  and  Shaffer  (1902)  found  that  ingestion  of  hydrochloric 
acid  increased  the  outgo  of  nitrogen,  sulphur  and  phosphorus,  with- 
out marked  change  in  the  proportions  between  these  elements. 

VonTabora  (1905)  studied  the  phosphorus  content  of  122  sam- 
ples of  gastric  juice  from  normal  and  pathological  subjects.  The 
phosphorus  content  of  the  stomach,  with  normal  or  greater  acidity, 
is  negligible,  following  a  phosphorus-free  test  meal,  the  maximum 
amounts  being  about  0.005  gm.  P205  in  10  c.c.  of  filtered  juice.  The 
calcium  content  was  about  0.002  gm.  in  100  c.c. 


200  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

The  author  concluded  that  the  phosphate  content  of  gastric 
juice  is  indirectly  proportional  to,  and  dependent  on,  the  degree  of 
acidity,  in  one  and  the  same  individual.  In  the  presence  of  hydro- 
chloric acid  all  phosphate  was  considered  to  be  present  in  the  diacid 
form ;  while  in  anacid  conditions  the  monacid  form  predominated. 

Fitz,  Alsberg  and  Henderson  (1907)  studied  urinary  phospho- 
rus elimination  in  rabbits  as  affected  by  the  ingestion  of  hydrochlo- 
ric acid.  The  introduction  of  acid  into  the  circulation  increased,  in 
four  rabbits,  the  outgo  of  phosphorus  in  the  urine.  This  increase 
was  followed  by  a  decrease,  and  then  after  further  introduction  of 
acid,  by  a  premortal  rise  in  phosphorus  outgo. 

This  increase  in  phosphorus  elimination,  under  the  influence  of 
acid  poisoning,  is  explained  by  the  conversion  of  di-  to  mono-phos- 
phate in  the  blood,  for  the  neutralization  of  the  acid,  and  the  res- 
toration of  the  normal  proportions  of  phosphorus  in  these  forms  by 
the  elimination  of  the  excess  of  mono-phosphate. 

Granstrom  (1908)  found  that  feeding  H3P04  to  rabbits  in- 
creased the  feces  calcium,  and  the  phosphorus  of  both  urine  and 
feces. 

Adler  (1909)  reports  a  study  of  the  effects  of  hydrochloric  acid 
and  calcium  phosphate  administration  on  the  excretion  of  calcium 
and  phosphorus  by  human  beings,  but  the  brevity  of  the  collection 
periods  renders  results  of  doubtful  significance. 

Wuertz  (1912)  found  that  the  addition  of  HC1  to  the  diet  of 
rabbits  did  not  affect  the  distribution  of  phosphorus  between  urine 
and  feces,  the  acid  being  so  rapidly  absorbed  that  its  ingestion  did 
not  increase  the  length  of  the  acid  portion  of  the  alimentary  tract, 
but  that  the  addition  of  calcium  carbonate  increased  the  alkalinity 
of  the  intestine,  and,  therefore,  greatly  increased  the  proportion  of 
the  phosphorus  excreted  in  the  feces. 

PHOSPHORUS  EXCRETION  AS  AFFECTED  BY  SALTS  OF  SODIUM  AND  POTASSIUM 

Sick  (1857)  found  that  the  ingestion  of  sodium  phosphate  in- 
creased urinary  phosphorus  by  more  than  the  added  amount,  with  a 
decrease  of  earth  phosphates,  and  increase  of  alkali  phosphates. 

C.  Ph.  Falck  (1872)  determined,  by  intravenous  injection  of  so- 
dium phosphate  into  dogs,  that  this  salt  was  promptly  eliminated  by 
the  kidneys,  the  principal  part  within  a  few  hours.  Doses  of  5.4 
to  10.2  gm.  by  intravenous  injection  (the  volume  of  the  solution  not 
stated)  caused  vomiting. 

Bunge  (1873)  found  that  potassium  phosphate,  as  well  as  the 
citrate  and  sulphate,  has  the  effect,  when  ingested  by  human  beings, 
to  cause  an  increase  in  the  elimination  of  sodium  salts. 


PHOSPHORUS  METABOLISM  201 

Bertram  (1878)  found  that  in  man  potassium  citrate  decreases 
urinary  excretion  of  phosphorus  but  little,  while  the  calcium  excre- 
tion is  greatly  diminished.  A  further  decrease  of  the  phosphoric 
acid  takes  place  if  in  addition  to  potassium  citrate,  calcium  carbon- 
ate is  also  given.  In  herbivora  Bertram  found  that  K2HP04  if  ad- 
ded, to  the  usual  diet  led  to  the  appearance  of  phosphorus  in  the 
urine. 

P.  A.  E.  Wagner  (1892)  demonstrated  the  diuretic  effect  of  di- 
sodium  phosphate  by  injecting  it  in  solution  into  the  jugular  vein  of 
rabbits.  The  urine  was  then  collected  at  15-minute  intervals.  The 
maximum  excretion  of  urine  was  during  the  second  15-minute  inter- 
val. 

Gerhardt  and  Schlesinger  (1899)  found  in  diabetes  that  inges- 
tion of  sodium  carbonate  reduced  urinary  phosphorus  excretion ;  but 
this  was  probably  by  virtue  of  its  alkalinity  alone,  and  not  by  virtue 
of  any  specific  action  of  sodium,  or  to  the  solubility  of  its  com- 
pounds. 

Desgrez  and  Guende  (1906)  studied  the  influence  of  phosphoric 
acid,  monosodium  phosphate  and  trisodium  phosphate  on  the  metab- 
olism of  the  guinea  pig.  Twenty-four  male  animals  were  used  in  four 
series  of  six  each.  The  basal  ration  was  consumed  ad  libitum, 
the  amounts  consumed  being  said  to  be  about  alike.  The  only 
phosphorus  metabolism  data  are  figures  representing  urinary  phos- 
phorus outgo.  The  urinary  nitrogen  was  increased  by  all  of  the 
phosphorus  compounds,  and  in  order  of  their  acidity,  that  is,  the 
neutral  phosphate  caused  the  least  increase,  the  acid  phosphate  the 
next,  and  the  phosphoric  acid  the  most. 

For  a  computation  by  Raoult's  formula  of  the  molecular  weight 
of  the  average  molecule  in  the  urine  in  these  experiments  see  Des- 
grez and  Posen  (1907). 

According  to  Loewi  (see  von  Noorden,  1907,  vol.  3,  p.  1079, 
1080;  German  ed.  vol.  2,  p.  685,  686)  von  Bunge  found  that  urinary 
phosphorus  excretion  is  not  influenced  by  sodium  salts,  but  that  po- 
tassium salts  cause  a  considerable  fall  in  urinary  phosphorus.  Loewi 
states  that  Bertram  cleared  up  the  question  as  to  the  method  of 
action  of  potassium  in  this  connection,  by  the  observation  (previous- 
ly mentioned)  that  the  exhibition  of  potassium  citrate  (er- 
roneously translated  "calcium  citrate,"  Eng.  ed.)  considerably  de- 
creases not  only  the  phosphoric  acid  but  also  the  calcium  in  the  urine 
(in  one  case  within  three  days  from  0.5  to  0.28  gram),  an  action 
which  is  not  shared  by  sodium  (Beckmann) .  The  cause  of  the  dif- 
ferent action  of  potassium  and  sodium  on  calcium  excretion  in  the 
urine  is,  according  to  Loewi,  who  credits  the  observation  to  Bert- 
ram, that  the  calcium  phosphates  found  in  the  body  fluids  are  insol- 


202 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


uble  in  potassium  carbonate,  and  hence  can  not  be  excreted  in  the 
urine,  while  they  are,  however,  soluble  in  the  corresponding  sodium 
compounds.  Loewi  states  that  the  excretion  of  magnesium  is  not 
affected  by  either  potassium  (Bertram)  or  sodium  (Beckmann). 

Hart,  McCollum  and  Humphrey  (1909),  in  their  study  of  phytin 
metabolism  with  a  dairy  cow,  found  that  the  potassium  of  the  feces 
varies  with  the  phosphorus  intake.  With  high  phosphorus  intake 
the  potassium  is  in  part  deflected  from  urine  to  feces,  while  with  a 
low  phosphorus  intake  the  potassium  is  eliminated  mostly  in  the 
urine. 

Von  Hoesslin  (1909),  in  experiments  with  dogs,  studied  the 
effects  of  sodium  chloride,  and  other  salts,  on  metabolism.  He  con- 
cluded that  the  addition  of  sodium  chloride  to  the  food  increased  the 
outgo  of  phosphorus,  especially  in  the  urine.  The  excretion  of  ter- 
tiary sodium  phosphate  by  the  kidneys  is  increased  by  water,  and 
still  more  by  the  sodium  chloride  intake.  The  phosphorus  outgo, 
especially  in  the  urine,  is  also  increased  by  overheating.  Phos- 
phates added  to  the  diet  caused  diuresis,  and  an  increase  in  the  per- 
centage content  of  the  urine  in  phosphorus. 

Oeri  (1909)  studied  phosphorus  metabolism  as  affected  by  in- 
gestion of  disodium  phosphate.  The  basal  ration  was  composed  of 
normal  foods  in  usual  combinations.  The  subjects  of  the  experi- 
ments were  a  woman,  aged  35  years,  weight  55  kg.,  and  the  author 
himself,  aged  25  years,  weight  93  kg.      The  numerical  data  follow. 

AVERAGE  DAILY  PHOSPHORUS  BALANCES  OF  MATURE  MAN  AND 

WOMAN  AS  AFFECTED  BY  INGESTION  OF  DISODIUM 

PHOSPHATE— Grams 


Periods  and 
days 

Intake 
P2O5 

Urine 
P2O5 

Feces 
P2O5 

Total 

excreted 

P2O5 

Urine, 

percent 

P2O5 

Feces, 

percent 

P2O5 

Bal- 
ance 
P2O5 

Diet 

Fore-period 
1-6 

Phosphate  fed 

7 
After-period 

8-12 
Phosphate  fed 

13 
After-period 

14 

6.45 

7.46 
5.45 
7.48 
5.45 

2.24 

2.62 
2.02 
2.64 
3.15 

2.50 

3.90 
3.07 
3.62 
2.83 

4.74 

6.52 
5.09 
6.26 
5.98 

47.3 

40.2 
39.7 
42.2 

52.7 

52.7 

59.8 
61.3 
57.8 
47.3 

+0.71 

+0.96 

+0.36 
+1.22 
-0.53 

Milk,  veal,  bouillon,  bread, 
potatoes,  apples,  sago, 
butter,  coffee,  jelly. 

Same  plus  disodium  phos- 
phate. 

Same  without  phosphate. 

Same  with  phosphate. 
Same  without  phosphate. 

Fore-period 
1-9 

Phosphate  fed 

10 
After-period 
11-17 

6.54 

8.57 
6.54 

3.62 

3.64 
3.39 

3.31 

4.06 

3.10 

6.93 

7.70 
6.49 

52.3 

47.0 
52.3 

47.7 

53.0 

47.7 

-0.39 

+0.87 
+0.06 

Milk,  veal,  bouillon,  bread, 
potatoes  apples,  pea  soup, 
butter,  coffee,  beer,  jelly. 
Same  plus  disodium  phos- 
phate. 
Same  without  phosphate. 

The  sodium  phosphate  was  without  marked  or  consistent  effectj 
on  the  partition  of  phosphorus  between  urine  and  feces. 


PHOSPHORUS  METABOLISM  203 

EFFECTS  OP  ■CALCIUM  AND  MAGNESIUM  ON  PHOSPHORUS  ELIMINATION 

Calcium  Carbonate.  Riesell  (1868)  found  that  the  ingestion 
(by  himself)  of  large  amounts  of  calcium  carbonate  (10  gm.  with 
each  meal  and  some  with  the  drink  )  decreased  the  urinary  phos- 
phorus to  about  one-half  the  normal,  but  during  four  days  of 
such  chalk  treatment  the  urinary  phosphorus  rose  nearly  to  the  nor- 
mal, apparently  through  the  absorption  of  the  calcium  phosphate 
formed  in  the  intestine.  Under  the  influence  of  the  chalk,  the  al- 
kali phosphates  of  the  urine  were  largely  replaced  by  alkaline  earth 
phosphates. 

Schetelig  (1880)  did  not  observe  a  decrease  of  urinary  phospho- 
rus from  taking  calcium  carbonate  for  2  days  in  3-gram  doses.  The 
amount  was  perhaps  insufficient. 

E.  Lehmann  (1882,  1894)  determined  that  the  alkaline-earth 
carbonates,  when  ingested,  have  the  effect  quantitatively  to  reduce 
the  urinary  phosphorus. 

Riidel  (1893a)  shows  that  the  administration  of  calcium  car- 
bonate may  cause  some  increase  in  the  urinary  calcium  of  children, 
but  not  by  any  means  in  proportion  to  the  intake. 

Strauss  (1896)  studied  the  effect  of  calcium  carbonate  inges- 
tion on  the  elimination  of  phosphorus,  uric  acid  and  purin  bases  in 
man.  Ingestion  of  calcium  carbonate  caused  a  decrease  in  urinary 
phosphorus,  which  did  not  disappear  immediately  after  the  with- 
drawal of  the  carbonate  from  the  food,  but  only  after  about  three 
days.  The  decrease  in  urinary  phosphorus  is  due  to  decrease  prin- 
cipally in  the  amount  of  monosodium  phosphate,  but  partially  to  de- 
crease in  disodium  phosphate,  thus  decreasing  the  urinary  acidity 
but  never  causing  alkalinity.  Uric  acid  and  purin  bases  were  not 
affected.      Urinary  calcium  was  slightly  increased. 

Herxheimer  (1897)  tested  the  effects  of  ingestion  of  calcium 
carbonate  baked  into  bread  in  the  amount  of  5  percent  of  the  same. 
The  carbonate  had  the  effect  to  decrease  the  urinary  phosphorus 
quite  markedly,  and  also  to  increase  the  feces  phosphorus,  though 
to  a  less  extent,  the  calcium  apparently  increasing  the  retention  of 
phosphorus,  though  phosphorus  in  the  food  was  not  determined. 
There  was  in  the  urine  a  greater  reduction  of  monophosphate  than 
of  diphosphate,  which  produced  in  the  latter  a  great  relative  in- 
crease. Herxheimer  states  that  18  grams  of  carbonate  baked  in 
the  bread  was  more  effective  than  30  grams  as  a  powder. 

Volhard  (1904)  found  that  great  quantities  of  calcium  carbon- 
ate in  the  food  did  not  seriously  affect  digestion  or  phosphorus 
metabolism. 


204  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

G.  Zuelzer  (1905)  states  that  calcium  affects  the  urinary  phos- 
phorus by  uniting  with  this  element  in  the  intestine  and  passing 
off  in  this  form  in  the  feces.  With  a  lime-poor  diet  the  feces  are 
poor  and  the  urine  is  rich  in  phosphorus. 

On  the  basis  of  work  by  Zerner  and  .  Ritter,  and  others, 
vonNoorden  and  Dapper  (vonNoorden  1907,  III,  946)  state  that 
even  after  the  ingestion  of  considerable  quantities  of  calcium  car- 
bonate the  urine  remains  slightly  acid,  since  the  calcium  is  excreted 
almost  entirely  by  way  of  the  intestine,  in  which  it  combines  with 
phosphoric  acid,  thus  restricting  its  absorption.  Thus  the  total 
phosphorus  of  the  urine  is  decreased,  and,  as  the  reaction  of  the 
urine  approaches  alkalinity,  the  proportion  of  disodium  to  mono- 
sodium  phosphates  is  modified  in  favor  of  the  former.  This  de- 
crease of  phosphates  involves  an  absolute  and  a  relative  decrease  of 
monosodium  phosphate,  a  substance  which  directly  promotes  the 
precipitation  of  uric  acid ;  while  the  relative  excess  of  disodium  phos- 
phate, which  is  capable  of  dissolving  uric  acid,  is  left  free  to  exert  a 
greater  effect. 

Kochmann  and  Petsch  (1911)  found,  in  metabolism  experi- 
ments with  dogs,  that  increasing  the  lime  of  the  diet,  the  phospho- 
rus remaining  constant,  increased  the  feces  phosphorus  and  de- 
creased the  urine  phosphorus.  When  a  calcium  equilibrium  had 
been  established  and  protein,  carbohydrates  and  fat  were  added  to 
the  ration,  the  calcium  intake  remaining  constant,  the  calcium  equi- 
librium was  disturbed,  and  lime  was  lost  from  the  bones. 

Bertram  (1878)  and  Renvall  (1904)  also  found  that  calcium 
carbonate  in  the  diet  served  to  deflect  phosphorus  from  urine  to 
feces. 

Various  Salts  of  Calcium  and  Magnesium.  Tereg  and  Arnold 
(1883)  reported  results  of  experiments  with  dogs  in  which  they 
compared  the  effects  .on  metabolism  of  the  ingestion  of  calcium  car- 
bonate and  primary,  secondary  and  tertiary  phosphates  as  supple- 
ments to  uniform  basal  rations. 

The  authors  interpret  the  negative  nitrogen  balance  in  the 
fourth  period  as  being  due  to  the  influence  of  the  primary  calcium 
phosphate  in  causing  increased  katabolism  of  body  pro- 
tein, and  cite  in  support  of  this  idea  the  coincident  loss  of  calcium 
and  phosphorus. 

We  would  suggest  that,  comparing  the  calcium  and  phosphorus 
balances  of  Periods  II,  III  and  IV,  the  smaller  the  proportion  of  cal- 
cium to  phosphorus  in  the  salt  supplement  the  less  is  the  storage,  or 
the  greater  the  loss,  of  both  calcium  and  phosphorus,  suggesting 
that  in  these  rations  the  lack  of  proportion  between  these  constit- 


PHOSPHORUS  METABOLISM 


205 


uents  restricts  the  usefulness  of  both.  The  great  increase  of  phos- 
phorus retention  caused  by  the  ingestion  of  lime  in  the  fifth  period 
shows  that  in  these  rations  the  lime  content  limited  the  deposition 
of  phosphorus. 


AVERAGE  DAILY  NITROGEN,  CALCIUM  AND  PHOSPHORUS  BALANCES 

WITH  A  DOG  ON  NORMAL  RATIONS  SUPPLEMENTED  BY 

INORGANIC  SALTS— Grams 


N 

CaO 

P2O5 

Period  and 
date 

Body  weight, 

initial  and 

final 

Food 
Urine 

Food 
Urine 

Food 
Urine 

P2O5:  N 
in  urine 

Rations 

Feces 

Feces 

Feces 

Balance 

Balance 

Balance 

I 

32000 

22.68 
17.76 
4.31 

0.780 

0.0455 

1.465 

5.040 
3.087 
3.003 

600  gm.  dog  biscuit 

Nov.  12-15 

32900 

+0.61 

-0.7305 

-1.049 

17:100 

II 

22.68 

5.140 

8.710 

600  gm.  dog-   biscuit; 

17.45 

0.1077 

3.539 

10  g-m.  Ca3  (P04)2 

4.90 

4.960 

4.728 

Nov.  18-21 

constant 

+0.33 

+0.0701 

+0.443 

17:100 

III 

22.68 

4.688 

9.350 

600  firm,  dog-  biscuit; 

17.30 

0.053 

4.496 

10  gm.  CaHP04 

5.10 

4.557 

4.810 

Nov.  29-Dec  2 

constant 

+0.28 

+0.078 

+0.044 

32:100 

IV 

32900 

22.68 

1.905 

9.255 

600  g-m.  dog-  biscuit; 

18.75 

0.0779 

5.895 

7.5  gm.  CaH4(P04>2 

4.83 

1.860 

3.853 

Dec  5-12 

32600 

-0.90 

-0.0354 

-0.493 

31:100 

V 

22.68 

6.380 
0.158 
4.310 

5.040 
1.336 

2.088 

600  gm.  dog  biscuit; 
10  gm.  chalk 

Dec  19-22 

+1.914 

+1.616 

VI 

33500 

22.58 

0.144 

2.802 

600    gm.  horse  flesh; 

16.45 

0.020 

2.532 

100  gm.  horse  fat 

3.90 

0.100 

0.237 

Dec  28-31 

33520 

+0.19 

+0.022 

+0.033 

19:100 

VII 

33520 

22.58 

4.052 
0.029 

7.112 
3.402 

600  gm.    horse    flesh; 

100  gm.  horse  fat; 

10  gm.  CaHP04 

Jan.  7-10 

33520 

VIII 

33520 

22.58 

0.894 
0.061 
0.740 

5.612 
4.572 
0.909 

600  gm.    horse    flesh; 
100  gm.  horse  fat; 

Jan.  13-16 

33430 

+0.093 

+0.131 

5  gm.  CaH4(P04>2 

Tereg  and  Arnold  compute  that  in  Period  IV  the  phosphorus 
loss  could  not  all  be  accounted  for  as  coming  from  the  loss  in  flesh, 
a  portion  apparently  coming  from  the  bones.  That  all  three  of 
these  phosphates  were  absorbed  is  not  to  be  questioned,  though 
the  authors  note  that  the  calcium  and  phosphorus  apparently  ab- 
sorbed, that  is,  missing  from  the  feces,  are  not  in  proportional 
amounts.     Either  these  salts  were  decomposed  in  the  intestine,  and 


206  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

the  parts  absorbed  independently  of  each  other,  or,  after  absorption, 
they  were  decomposed  in  the  blood,  and  the  parts  eliminated  in  un- 
like portions  through  the  intestines  and  the  kidneys. 

Of  the  primary  salt  (Period  IV)  nearly  twice  as  much  calcium 
and  phosphorus  were  eliminated  in  the  urine  as  of  the  secondary 
salt.  In  periods  VI,  VII  and  VIII  where  the  basal  ration  was  meat, 
the  primary  salt  caused  a  much  greater  increase  of  the  calcium 
and  phosphorus  of  the  urine  than  did  the  secondary  salt,  and  in  in- 
jection experiments  the  authors  noted  similar  increase  of  the  cal- 
cium and  phosphorus  of  the  urine.  It  is  their  opinion  therefore 
that  the  primary  salt  is  absorbed  as  such  from  the  intestine. 

Steel  and  Gies  (1907a)  determined  the  effects  of  the  addition 
of  bone  ash  to  a  diet  of  prepared  meat,  cracker  meal,  lard  and  water, 
on  the  urinary  excretion  of  calcium  and  phosphorus  by  dogs.  It 
was  desired  to  use  bone  ash  in  the  ration  to  give  bulk  and  body  to 
the  feces. 

Bone  ash  seems  not  to  alter  the  urinary  elimination  of  cal- 
cium, but  a  decrease  of  urinary  phosphorus  from  0.460 
to  0.244  gm.  P205  in  21  days,  during  the  feeding  of  bone  ash,  shows 
that  the  use  of  bone  ash  for  the  purpose  intended  would  be  justifi- 
able only  in  such  experimental  work  as  would  be  unaffected  by  a  de- 
crease of  phosphorus  absorption  or  change  in  path  of  outgo. 

Lothrop  (1909)  investigated  the  effect  of  bone  ash  in  the  diet 
on  the  metabolism  of  the  dog.  The  ingestion  of  bone  ash  decreased 
the  urinary  phosphorus. 

Hart,  McCollum  and  Humphrey  (1909)  found,  in  their  study  of 
phytin  metabolism  with  a  milch  cow,  that  calcium,  magnesium  and 
phosphorus  were  excreted  principally  in  the  feces,  and  that  a  low 
phosphorus  intake  served  to  increase  the  urinary  elimination  of  cal- 
cium. 

R.  Berg  (1910a)  conducted  balance  experiments  on  himself, 
using  a  basal  ration  to  which  he  added,  in  different  periods,  various 
organic  and  inorganic  phosphorus  compounds.  Tricalcic  phosphate 
he  found  excreted  largely  as  such  in  the  feces.  The  dicalcic  phos- 
phate was  excreted  in  part  as  tricalcic  phosphate  and  magnesium 
ammonium  phosphate.  Hypophosphites  were  almost  all  excreted 
unchanged.  Berg  considers  that  the  best  source  of  calcium  is  green 
vegetables  prepared  so  as  to  retain  their  salts. 


PHOSPHORUS  METABOLISM  207 

De  Jager  (1910)  submits  the  following  data  showing  the  effects 
of  calcium  salts  on  urinary  phosphorus  in  man. 

URINARY  PHOSPHORUS  AS  AFFECTED  BY  CALCIUM  SALTS— Grams 

Days  Periods                                                                                               Ave.  Daily  PaOs 

4  Preliminary .2.462 

10         4  gm.  Ca-sulphate  daily 2.344 

16         Interval    2.489 

5  7.5  gm.  Ca-lactate  daily 2.087 

18         Interval    2.199 

10  6  gm.  Ca-sulphate  daily 1.767 

14         After  period    1.999 

The  slight  decrease  of  urinary  phosphorus  produced  by  the  cal- 
cium sulphate  is  doubtless  due  to  the  relative  insolubility  of  this 
salt.  The  lactate,  with  its  weak,  oxidizable  acid  radical,  yields 
much  more  of  its  calcium  free  for  combination  and  excretion  with 
phosphorus  in  the  feces. 

A.  R.  Rose  (1912a),  in  his  phytin  metabolism  study  with  a  cow, 
found  that  the  ingestion  of  calcium  and  phosphorus  as  the  phytate 
deflected  a  part  of  both  the  calcium  and  magnesium  from  urine  to 
feces,  the  calcium  added  as  the  phytate  also  being  eliminated  by  the 
feces.  With  decreasing  phosphorus  in  the  food  there  was  an  in- 
crease of  urinary  calcium. 

Gregersen  (1911)  showed  with  rats  that  calcium  and  magnes- 
ium tend  to  deflect  phosphorus  excretion  into  the  feces,  either  when 
the  phosphorus  excreted  is  contained  in  the  food,  or  when  it  is  met- 
abolized phosphorus  from  the  body.  Gregersen  also  found  that 
this  effect  was  not  affected  by  such  acid-saturating  salts  as  sodium 
bicarbonate. 

In  experiments  with  swine,  Forbes,  Beegle,  Fritz  and 
Mensching  (1914)  found  that  in  common  foods  magnesium  tends  to 
deflect  the  phosphorus  excretion  from  urine  to  feces,  and  excess- 
ive phosphorus  content  of  the  ration  seems  to  limit  the  ab- 
sorption of  magnesium.  With  an  average  daily  intake  of  2.17  gm. 
magnesium  and  5.40  gm.  phosphorus  there  was  storage  of  magne- 
sium, but  with  an  intake  of  9.28  gm.  magnesium  and  20.71  gm. 
phosphorus  there  was  loss  of  magnesium,  apparently  combined  with 
phosphorus,  through  the  feces. 

EFFECTS  OF  DIET  AND  SPECIES  ON  PHOSPHORUS  ELIMINATION 

The  bearing  of  species,  especially  as  determining  prevailing 
diet,  on  the  paths  of  elimination  of  phosphorus  is  marked.  The 
herbivora  normally  consume  a  diet  which  is  comparatively  poor  in 
phosphorus  and  rich  in  alkalis  and  alkaline  earths.  The  omnivora 
and  carnivora,  on  the  other  hand,,  live  on  a  diet  which  is  so  much 
richer  in  phosphorus  and  poorer  in  mineral  bases  that  a  different 
and  much  more    extensive    acid-neutralizing    capacity    has    been 


208  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

evolved.  Thus,  the  herbivora  excrete  their  mineral  acids  in  com- 
bination with  fixed  alkalis,  while  the  omnivora  and  carnivora  are 
able  to  excrete  the  same  largely  combined  with  ammonia.  During 
the  suckling  period,  however,  the  excretion  of  phosphates  is  much 
the  same  in  both  classes  of  animals.  A  few  of  the  many  papers 
showing  the  different  methods  of  elimination  of  phosphorus  in  herb- 
ivora and  other  animals  are  mentioned  in  brief. 

Bischoff  (1867)  found  that  in  dogs  phosphorus  is  excreted  to 
the  extent  of  about  12-thirteenths  in  the  urine,  mostly  combined 
with  the  alkalis,  while  the  remainder  leaves  the  body  in  the  feces 
combined  principally  with  calcium,  iron  and  magnesium. 

Bertram  (1878)  concluded  that  the  absence  of  phosphorus  from 
the  urine  of  herbivora  was  due  to  the  richness  of  the  food  in  vege- 
table alkali  compounds  and  calcium  salts,  alkaline  liquids  containing 
carbonates  having  no  solvent  power  for  calcium  phosphate,  though 
they  do  dissolve  magnesium  phosphate. 

F.  Miiller  (1884)  concluded  from  analyses  of  meconium  and  of 
feces  from  diets  of  meat  that  the  dog  excretes  its  lime  mostly 
through  the  feces,  while  magnesium,  and  to  a  greater  extent  still, 
phosphorus,  is  excreted  chiefly  through  the  urine.  The  content 
of  the  feces  in  the  alkalis  varies  indirectly  with  the  length  of  stay  in 
the  intestine. 

Weiske  (1872b)  analyzed  the  urine  of  two  goats,  one  receiving 
milk  alone,  ad  libitum,  and  the  other  green  clover  and  turnip  leaves. 
The  ash  analyses  are  as  follows : 

Vegetable  diet  Milk  diet 

K2O 34.91 42.83 

Na20 22.48 14.05 

CaO 0.77 „ 0. 98 

MgrO 3.28 0.61 

Fe203 trace trace 

CO2 10.40 none 

Si02 „ 0.59 none 

S03 16.89 3.02 

P2O5 trace 22.22 

CI 13.35 20.67 


102.67  104.38 

O  out  for  CI 3.01  4.66 

99.67  99.72 

On  a  milk  diet  the. goat  excretes  much  phosphorus  in  the  urine; 
on  this  vegetable  diet  no  phosphorus  was  excreted  in  the  urine. 

Jordan  (1885-6)  found  in  the  urine  of  a  sheep  nearly  half  of  the 
total  potassium  outgo,  but  no  phosphorus. 

E.  Wolff  (1886)  reports  determinations  of  the  phosphorus  of 
clover  hay,  and  of  the  feces  resulting  from  its  being  fed  to  horses. 
The  feces  phosphorus  equalled  the  food  phosphorus.  With  a  ration 
of  oats,  straw  and  meadow  hay  the  feces  phosphorus  was  98.9  per- 
cent of  the  food  phosphorus. 


PHOSPHORUS  METABOLISM  209 

Grundzach  (1892)  determined  the  ash  constituents  of  normal 
human  feces,  and  found  in  the  ash  29.25  percent  CaO  and  13.76  per- 
cent P205. 

Paton,  Dunlop  and  Aitchison  (1899-1900)  made  a  general  study 
of  the  paths  of  elimination  of  phosphorus  compounds  by  animals, 
and  their  principal  findings  are  as  follows : 

In  dogs,  on  a  vegetable  diet,  a  large  proportion  of  the  phospho- 
rus of  the  food  is  not  excreted  in  the  urine. 

In  dogs,  on  a  diet  of  dog's  biscuit  and  milk,  a  large  proportion 
of  the  phosphorus  injected  subcutaneously  as  sodium  phosphate  is 
not  excreted  in  the  urine  (no  food  or  feces  data) . 

During  lactation  there  is  a  diminished  urinary  excretion  of 
phosphorus  by  the  dog,  and  a  diminished  feces  excretion  of  phos- 
phorus by  the  goat. 

In  the  goat  none  of  the  phosphoric  acid  injected  subcutaneous- 
ly as  sodium  phosphate,  or  formed  in  the  body,  or  contained  in  the 
food,  is  excreted  in  the  urine. 

The  administration  of  calcium  glycerophosphate  by  the  mouth 
causes  no  increased  excretion  of  phosphorus  in  the  urine  of  the  dog, 
or  in  the  urine  or  milk  of  the  goat. 

Rumpf  and  Schumm  (1900)  made  urinary  phosphorus  esti- 
mations in  a  human  metabolism  experiment  with  a  vegetable  diet. 
The  subject,  weighing  62.5  kg.  at  the  beginning  of  the  8-day  experi- 
ment, gained  1.7  kg.  during  this  time  on  a  ration  supplying  3431 
calories  and  73.88  gm.  protein  daily.  The  urinary  nitrogen  was  6.91 
gin.,  and  urinary  phosphorus  (P205)  1.38-2.43  gm.  per  day. 

Bergmann  (1901)  studied  phosphorus  elimination  in  the  dog 
and  the  sheep.  Sodium  phosphate  injected  subcutaneously  into  a 
dog  receiving  a  ration  of  meat  and  bread  was  eliminated  in  the 
urine.  The  same  result  was  obtained  when,  in  addition  to  the  phos- 
phate injection,  calcium  carbonate  was  added  to  the  diet. 

With  a  wether,  on  a  diet  of  hay  and  oats,  sodium  phosphate  was 
injected  subcutaneously,  and  all  of  it  was  excreted  by  the  intestine, 
the  urinary  phosphorus  remaining  unweighable. 

Glycerophosphoric  acid,  likewise,  when  subcutaneously  inject- 
ed, was  excreted  by  the  dog  in  the  urine,  and  by  the  wether  in  the 
feces,  as  inorganic  phosphoric  acid.  Bergmann  also  notes  the  well- 
known  fact  that,  while  dogs  normally  excrete  P  mostly  in  the  urine, 
if  Ca  is  abundant  in  the  food  much  P  is  excreted  with  it  in  the  feces. 

Tangl  (1901)  found  in  metabolism  experiments  with  horses,  on 
hay  and  oats,  that  more  than  half  of  the  calcium,  about  three- 
fourths  of  the  magnesium,  and  nearly  all  of  the  phosphorus  of  the 


210  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

food  is  eliminated  in  the  feces.  Tangl  concluded  that  a  horse  was 
able  to  obtain  its  calcium  needs  from  the  same  low-calcium  fodder 
which  with  cattle  may  produce  malnutrition  of  the  bones.  See  also 
W.  Camerer,  Sr.  (1904) ;  composition  of  human  urine  as  affected  by 
animal  and  vegetable  diet,  disease,  etc. 

Gouin  and  Andouard  (1907)  conducted  experiments  showing 
that  a  calf  receiving  milk  alone  excreted  83.33  percent  of  the  food 
phosphorus  in  the  urine.  An  addition  of  powdered  bone  to  the  food 
did  not  alter  the  percent  of  the  total  phosphorus  excreted  in  the 
urine. 

Another  calf  received  55.6  gm.  P205  in  milk  and  vegetable  foods, 
and  excreted  13.74  percent  of  this  amount  in  the  urine;  when  29.22 
gm.  more  P205  was  added  in  the  form  of  powdered  bone  the  amount, 
of  phosphoric  acid  in  the  urine  was  increased  by  2.36  gm.,  and  the 
percent  of  the  total  phosphoric  acid  of  the  food  found  in  the  urine: 
was  reduced  to  11.79. 

A  third  calf  received  no  milk  in  the  food.  The  urine  contained 
1.15  gm.  phosphoric  acid,  which  was  7.85  percent  of  the  total  ex- 
cretion. When  33.5  gm.  P205  was  added  to  the  diet  in  the  form  of 
bone,  the  quantity  in  the  urine  increased  to  6.35  gm.,  and  the  per- 
cent of  the  total  excretion  to  20.92. 

Another  calf  passed  no  phosphorus  in  the  urine.  Powdered 
bone  was  added  to  the  diet  in  the  amount  of  18.31  gm.  daily  for  7 
weeks.  The  storage  of  phosphoric  acid  was  17.5  gm.  per  kilogram, 
of  increase  in  live  weight. 

These  data  show  that  bone  phosphate  may  be  retained  by- 
calves,  and  that  the  vegetable  foods  used  tended  to  deflect  the  ex- 
cretion of  the  absorbed  phosphorus  of  the  bone  meal  from  urine  to 
feces.  The  authors  (G.  and  A.)  referred  these  results  to  differen- 
ces in  digestibility  of  the  phosphorus  in  the  different  forms. 

Magnus-Levy  (vonNoorden  1907,  I,  p.  41)  speaks  as  follows 
regarding  the  effects  of  species  and  diet  on  phosphate  elimination : 

"The  complete  removal  of  the  phosphoric  acid  from  the  urine 
of  man  and  of  dogs,  as  in  the  case  of  herbivora,  cannot  be  effected. 
Acid  sodium  phosphate,  when  subcutaneously  injected  into  sheep,  is 
excreted  in  the  feces,  and  in  dogs  appears  almost  entirely  in  the 
urine,  even  when  the  intestine  is  overladen  with  calcium  salts 
(Bergmann) . 

"In  dogs  the  separation  of  phosphoric  acid  by  the  kidneys  is  not 
affected  by  the  addition  of  alkali  if  the  food  is  not  rich  in  calcium 
(Beckmann) ;  the  phosphoric  acid  salts  of  the  sodium  and  cal- 
cium of   the    food   then   pass    out    almost    entirely    by   way    of: 


PHOSPHORUS  METABOLISM 


211 


the  kidneys  (Markuse,  Leipziger,  Zadik) .  It  is  only  when  an  ex- 
cess of  calcium  and  alkali  are  together  present  in  the  food  that  the 
phosphoric  acid  usually  excreted  by  the  kidneys  is  conducted  from 
and  to  the  intestine." 

Gouin  and  Andouard  (1908)  attempted  to  answer  the  question 
as  to  why  adult  cattle  do  not  secrete  phosphorus  in  the  urine.  In 
the  light  of  current  understanding,  however,  their  conclusions  as  to 
kidney  resistance  to  phosphates  are  no  longer  significant. 

Oeri  (1909)  found  that  with  mature  human  beings  the  excre- 
tion of  calcium  and  phosphorus  by  the  intestine  is  increased  (1)  by 
diets,  which,  like  milk,  contain  considerable  inorganic  phosphorus 
and  calcium,  (2)  by  a  diet  rich  in  phosphorus,  when  calcium  is 
separately  administered,  and  (3)  by  a  calcium-rich  diet,  when 
inorganic  phosphorus  is  superposed,  that  is,  whenever  phosphorus 
and  calcium  are  associated  within  the  body. 

Stutzer  (1910)  submits  analyses  of  the  urine  and  feces  of  the 
common  farm  animals,  showing  the  content  of  mineral  elements  in 
these  excreta.  The  cows  used  were  animals  giving  12-15  liters  of 
milk  on  medium  rations  of  green  and  dry  fodder.  The  sheep  received 
hay  only.  To  the  pigs  were  fed  potatoes  with  green  fodder,  but  no 
milk.  The  specific  gravity  of  the  urine  was  as  follows :  Sheep  1.038 ; 
horse,  1.035;  cow,  1.037;  pigs,  1.020.  There  was  much  similarity  in 
the  methods  of  elimination  of  the  several  mineral  elements  by  these 
animals.  The  influence  of  species  on  the  paths  of  excretion  could 
only  be  determined  by  feeding  the  animals  to  be  compared  all  on  the 
same  ration. 


ANALYSES  OF  FRESH  URINE  AND  FECES— Parts  per  1000 


Urine 

Fe 

ces 

Sheep 

Horse 

Cow 

'  Pig 

Sheep 

Horse 

Cow 

Pig 

Water    

903 

926 

923 

966 

680 

750 

835 

800 

Organic  material    . 

70 

47 

57 

23 

295 

230 

150 

160 

Nitrogen 

(a)    Total   amount 

15.8 

15.2 

15.0 

6.4 

6.2 

5.6 

5.9 

6  0 

(b)  Easily  soluble 

15.8 

15.2 

15.0 

6.4 

0.5 

0.5 

0.6 

0.8 

Phosphoric  acid 

(a)    Total   amount 

1.3 

0.05 

1.5 

1.6 

3.0 

3.0 

2.8 

6  0 

(b)  Easily  soluble 

1.3 

0.05 

1.5 

1.6 

0  5 

18.5 

16.5 

15.5 

.8.0- 

1,7 

3.3 

1.4 

5  0 

1.8 

3.2 

0.3 

0.1 

4.0 

2.3 

2.4 

0.5 

Magnesium     

2.5 

2.4 

0.1 

0.8 

2.4 

1.0 

1.8 

0  2 

Sulphuric   acid    . . . 

-     1.0 

1.6 

0.3 

2.7 

1.4 

0.5 

1.2 

0  6 

3.8 

3.0 

1.0 

1.0 

1.0 

0.1 

0.1 

0.1 

Summary.  The  ingestion  of  acids  or  acid  salts,  or  acid-form- 
ation in  the  body,  increases  urinary  calcium  and  phosphorus,  and  in- 
creases the  proportion  of  acid  phosphates  to  total  phosphates  in  the 
urine. 


212  OHIO  EXPEKIMENT  STATION:  TECHNICAL  BUL.  5 

The  alkaline  earths  decrease  urinary  phosphorus  elimination 
by  uniting  with  phosphorus  in  the  intestine  to  form  difficultly  sol- 
uble salts,  thus  hindering  phosphorus  absorption. 

Calcium  carbonate  causes  a  considerable  replacement  of  alkali 
phosphates  by  alkaline  earth  phosphates  in  the  urine.  The  de- 
crease of  alkali  phosphates  is  more  largely  mono-  than  di-sodium 
phosphate. 

Sodium  phosphate  is  promptly  eliminated  by  the  kidneys  and 
has  a  diuretic  effect.  Sodium  chloride  also  has  some  tendency  to 
increase  the  urinary  phosphorus.  Sodium  carbonate  may  reduce 
urinary  phosphorus  excretion  when  this  is  due  in  part  to  acidosis. 

Potassium  phosphate,  as  well  as  other  potassium  salts,  increas- 
es the  excretion  of  sodium  salts.  Potassium  phosphate  added  to 
the  usual  diet  of  herbivora,  leads  to  the  appearance  of  phosphorus 
in  the  urine.  In  man,  potassium  citrate  causes  a  fall  of  calcium 
phosphate  in  the  urine  by  virtue  of  the  insolubility  of  calcium  phos- 
phates in  potassium  carbonate.      Sodium  salts  have  not  this  effect. 

Calcium  phosphates  affect  phosphorus  outgo  in  the  urine  in  ac- 
cord with  their  acidity;  the  more  acid  the  salt,  the  greater  is  the 
urinary  excretion  of  calcium  and  phosphorus.  The  ingestion  of 
bone  ash  decreases  urinary  phosphorus,  probably  through  hindering 
phosphorus  absorption. 

Calcium  lactate,  like  the  carbonate,  may  decrease  urinary  phos- 
phorus. Calcium  sulphate  appears  to  decrease  urinary  phosphorus 
to  a  very  slight  degree. 

The  phosphorus  of  the  diet  of  carnivora  is  eliminated  principal- 
ly in  combination  with  alkalis  in  the  urine.  The  remainder  is 
eliminated  in  combination  with  iron,  magnesium  and  calcium  in  the 
feces.  Carnivora  excrete  their  magnesium  more  largely  than  their 
calcium  in  the  urine. 

Sodium  phosphate,  injected  subcutaneously  in  the  dog,  is  ex- 
creted in  the  urine  even  if  calcium  carbonate  be  added  in  quantity 
to  the  food. 

Calcium  added  to  the  food  of  carnivora  increases  the  feces  phos- 
phorus ;  and  on  a  vegetable  diet  carnivora  excrete  a  large  part  of  the 
phosphorus  in  the  feces. 

Herbivora,  in  accord  with  the  high  calcium  content  of  the  diet, 
excrete  almost  all  of  their  phosphorus  in  the  feces,  the  urine  usually 
being  practically  free  from  phosphorus.  Subcutaneous  injections 
of  sodium  phosphate  are  excreted  in  the  feces. 

During  starvation,  or  on  an  animal  diet,  such  for  instance  as 
milk,  herbivora  excrete  much  calcium  and  phosphorus  in  the  urine. 


PHOSPHORUS  METABOLISM  213 

During  lactation  there  is,  with  carnivora,  a  decrease  of  urinary- 
phosphorus  ;  with  herbivora  there  is  a  decrease  of  feces  phosphorus. 

Human  beings  excrete  their  phosphorus  according  to  the  same 
laws  as  carnivora  and  herbivora,  the  path  of  outgo  varying  with  the 
diet.  The  prevalence  of  meat  and  cereals,  which  have  acid  ash,  in 
ordinary  mixed  diets,  deflects  the  larger  part  of  the  phosphorus  into 
the  urine.  Vegetable  diet  or  calcium  carbonate  increases  the  feces 
phosphorus,  but  the  complete  deflection  of  phosphorus  from  the 
urine  to  the  feces  has  not  been  accomplished. 

On  a  milk  diet  three-fourths  of  the  phosphorus  is  excreted  in 
the  urine  by  infants,  and  nearly  the  whole  of  it  by  calves. 

The  alkaline  earths  deflect  metabolic  phosphorus  as  well  as  food 
phosphorus  into  the  feces,  and  fasting  feces  contain  considerable 
amounts  of  calcium  and  magnesium  phosphate. 

The  balance  of  magnesium  may  be  changed  from  positive  to 
negative  in  swine  by  excessive  ingestion  of  phosphorus  in  common 
foods,  the  loss  occurring  through  the  feces. 

RELATIVE  METABOLISM  OF  PHOSPHORUS  AND  OTHER 

ELEMENTS 

There  is,  without  doubt,  a  certain  interdependence  of  all  of  the 
constituents  of  the  body,  in  their  metabolism ;  for  instance,  if  nitro- 
gen is  lacking  in  the  ration  the  metabolism  of  all  other  elements 
which  are  used  with  nitrogen  will  be  affected  in  some  measure,  the 
degree  depending  on  the  extent  and  duration  of  the  nitrogen  de- 
ficiency. 

The  less  the  available  supply  of  any  nutrient,  compared  with 
the  demand  for  the  same,  the  more  definitely  does  it  become  the  lim- 
iting factor  in  production,  and  the  more  closely  does  its  metabolism 
measure  that  of  the  body  as  a  whole.  Any  element,  therefore, 
might  become  the  limiting  factor  in  metabolism ;  thus,  on  a  ration 
which  is  practically  iron-free,  the  total  growth  might  vary  directly 
with  the  iron  content  of  the  ration. 

In  view  of  their  distribution  and  relationships  in  the  tissues  it 
might  be  supposed  that  nitrogen,  sulphur,  calcium  and  phosphorus 
would  ordinarily  be  found  somewhat  closely  interdependent  in  met- 
abolism; and  since  phosphorus,  as  a  universal  cell  constituent,  is 
concerned  in  all  growth  and  production,  it  would  be  assumed  that 
this  element  would  be  found  quantitatively  related  especially  to 
those  other  elements  whose  metabolism  is  dominant  during  the  par- 
ticular kind  of  growth  or  production  under  observation ;  thus  for  the 
maintenance  and  repair  of  the  mature  animal,  nutrients  are  used  in 


214  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

certain  rather  definite  proportions,  one  to  another;  during  normal 
growth  the  proportions  would  be  somewhat  different,  and  it  is  quite 
conceivable  that  pregnancy,  lactation,  egg-laying  and  wool-growing 
should  each  have  some  special  requirements  for  nutrients  which 
would  affect  their  relative  quantitative  metabolism;  further,  con- 
ditions may  be  favorable  for  the  growth  of  bone,  without  providing 
liberally  for  other  increase.  In  each  of  these  phases  of  normal 
metabolism  phosphorus  is  involved,  and  its  quantitative  relations  to 
other  elements  have  been  the  subject  of  study  and  record,  the  ob- 
ject being,  ordinarily,  the  determination  of  the  states  and  conditions 
of  nutrition  as  made  manifest  by  the  quantitative  and  qualitative 
estimation  of  the  constituents  which  have  been  concerned.  A  meth- 
od of  frequent  use  is,  for  instance,  the  estimation  of  relative  metab- 
olism of  bone  and  soft  parts  by  the  relation  of  the  details  of  the  in- 
come and  outgo  to  the  composition  of  these  tissues. 

We  have  as  the  several  factors  of  such  considerations  first, 
two  possible  sources  of  the  constituents  involved;  the  nutrients  of 
the  food,  and  products  of  tissue  katabolism;  second,  two  possible 
methods  of  disposition  of  these  constituents,  namely,  retention  and 
outgo.  Usually  our  observations  are  confined  to  the  determina- 
tion of  final  or  net  balance,  the  relative  amounts  of  the  constituents 
retained  or  lost  indicating  the  identity  of  the  parts  involved. 

Some  limitations  of  the  value  of  this  method  of  study  are  that 
we  do  not  know  that  all  organs  are  katabolized  en  masse,  nor  that 
this  process  is  not  to  any  extent  partial  or  selective;  and  further, 
we  know  very  little  of  the  resynthesis  of  katabolized  nutrients. 
Participation  of  products  of  tissue  katabolism  in  synthetic  or  reten- 
tion processes  may  sometimes  be  inferred  from  the  outgo  of  accom- 
panying products  which  are  not  retained ;  thus  purin  nitrogen  elim- 
ination may  signify  the  katabolism  of  nucleins  under  circumstances 
where  the  subsequent  retention  of  the  accompanying  phosphorus 
and  non-purin  nitrogen  would  mask  this  process. 

In  spite  of  the  uncertainties  attending  these  attempts  to  judge 
of  the  origin  of  katabolized  nutrients  from  the  composition  of  the 
excreta,  the  method  has  its  uses,  though  in  times  past  it  has  been 
overworked. 

The  supplementary  capacities  of  the  intestine  as  a  means  of 
elimination  of  katabolized  constituents  require  that  the  feces  as  well 
as  the  urine  be  involved  in  such  studies.  Conclusions  resting  on  urine 
analyses  alone  must  be  considered  as  open  to  question. 

Bischoff  (1867)  concluded  that  nitrogen  and  phosphorus  ex- 
cretion rise  and  fall  together,  except  in  fasting ;  the  phosphorus  ex- 
cretion then  being  relatively    greater,    the    increase    being    due, 


PHOSPHORUS  METABOLISM  215 

according  to  Bischoff ,  to  a  loss  of  phosphates  from  the  plasma,  with- 
out corresponding  protein  metabolism.  This  extra  phosphorus  out- 
go in  fast  is  now  often  ascribed  to  bone  katabolism. 

J.  Forster  (1873b)  published  urinary  nitrogen  and  phosphorus 
data  from  a  man  on  a  meat  diet  which  show,  in  considerable  meas- 
ure, at  least  a  temporary  independence  of  nitrogen  and  phosphorus 
metabolism.      The  figures  are  given  below. 

EXCRETION  OF  NITROGEN  AND  PHOSPHORUS  IN  THE  URINE  OF  A 
HEALTHY  ADULT  ON  A  MEAT  DIET  DURING  TWENTY-FOUR 

HOURS— Grams 


Hours  during1  the  day 

N  , 

P2O5 

10  A.  M.— 1  P.  M 

2.74 
3.51 
3.36 
3.36 
2.52 
2.56 

0.76 

2  P.  M.— 5  P.  M 

0.62 

6  P.  M.— 9  P.  M. 

0.42 

10  P.  M.—  1  A.  1VL 

0.41 

2  A.  M.— 5  A.  M 

0.32 

6  A.  M.— 9  A.  M 

0.29 

A  healthy  adult  ingested  at  9  A.  M.  500  gm.  of  finely  chopped 
flesh,  containing  18.04  gm.  N  and  48.3  gm.  fat,  after  having  fasted 
since  the  preceding  noonday  meal.  He  then  drank  water  but  took 
no  other  food  for  24  hours.  During  this  time  he  excreted  nitrogen 
just  about  equal  to  the  total  present  in  the  500  grams  of  meat;  and 
of  phosphorus,  we  compute,  3  times  as  much  as  was  in  the  meat. 
These  facts,  taken  in  connection  with  the  rates  of  excretion  as 
shown  by  the  above  figures,  exhibit  the  truth  that  there  is  no  close 
relation  between  urinary  nitrogen  and  phosphorus  excretion.  The 
large  phosphorus  excretion  and  the  steady  decline  in  the  rate  of 
elimination  show  the  influence  of  the  previous  diet. 

W.  Zuelzer  (1876)  published  extensive  papers  on  the  propor- 
tions of  nitrogen  and  phosphorus  in  the  urine  of  human  beings  and 
of  dogs  in  various  states  of  nutrition.  In  the  absence  of  food  and 
feces  data  the  significance  of  the  conclusions  is  limited,  but  certain 
notes  of  interest  are  recorded. 

With  a  dog,  the  P205  of  the  urine  from  a  fat-free  flesh  diet  was 
12.8  percent,  during  fast  8.7  percent,  while  receiving  potatoes  27.1 
percent,  and  while  receiving  rye  bread  25.3  percent  of  the  amount  of 
the  nitrogen. 

The  chief  meal  of  the  day  exerts  an  influence  on  the  nitrogen 
and  phosphorus  of  the  urine  of  the  hours  immediately  following, 
while  this  influence  diminishes  with  lapse  of  time  thereafter. 

In  a  table  of  ratios  of  phosphorus  to  nitrogen  in  the  urine  at 
different  ages,  from  infancy  to  old  age,  it  is  shown  that  the  propor- 
tion of  phosphorus  to  nitrogen  is  greatest  in  infancy,  and  decreases 


216  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

rapidly  during  the  period  of  growth.  There  appears  to  be  little 
change  during  the  middle  years  of  life,  but  there  is  evidence  of  a 
slight  increase  of  phosphorus  in  proportion  to  nitrogen  in  old  age. 
These  figures  must  depend  largely  on  the  prevailing  diet  at  the  var- 
ious ages.  Zuelzer  was  of  the  opinion  that  increased  phosphorus 
excretion  signified  loss  from  the  nervous  system,  an  idea  now  long 
since  abandoned. 

C.  Voit  (1881)  submitted  the  following  figures  for  the  propor- 
tions of  nitrogen  and  phosphorus  in  dog  urine  under  different  con- 
ditions : 

N:P205  on  meat  diet  8.1:1  Bischoff 

N  :P205  during  fast  6.4 :1 

N:P205,  diet  of  N-poor  bread  3.8:1 

Feder  (1881)  attempted  to  determine  a  normal  ratio  of  P,  S 
and  N  in  the  urine  during  fast,  and  feeding  on  various  diets.  This 
he  found  it  impossible  to  do.  He  also  showed  that  the  hourly  vari- 
ations in  the  phosphorus  outgo,  and  in  the  proportion  of  nitrogen  to 
phosphorus  in  the  outgo,  could  not  be  due  directly  to  nerve  katabol- 
ism,  a  fact  which  becomes  obvious  when  we  consider  the  amounts  of 
these  variations  in  connection  with  the  phosphorus  content  of  the 
nervous  system. 

W.  Zuelzer  (1881a)  determined  the  proportion  of  N:P205  in  the 
muscular  system  as  100:15,  in  nerve  substance  as  100:  45  and  in 
blood  as  100 :4.  He  made  much  of  the  idea  (1881b)  of  determining 
the  source  of  the  nitrogen  and  phosphorus  of  the  urine  by  their 
proportionate  amounts,  but  ignored  the  feces.  Zuelzer  (1881a) 
stated  that  a  large  relative  amount  of  phosphorus  as  compared  with 
nitrogen  in  the  urine,  or  a  high  content  of  glycerophosphoric  acid, 
indicates  nerve  katabolism. 

Politis  (1884)  considered  the  proportions  of  nitrogen  and  phos- 
phorus in  the  urine  of  dogs  receiving  brain  in  the  food,  and  also 
the  hourly  variation,  during  two  days,  of  the  nitrogen  and  phospho- 
rus of  the  urine.  He  reached  the  conclusion  that  urinary  nitrogen 
and  phosphorus  data  alone  were  insufficient  to  demonstrate  the 
breaking  down  of  brain  substance.  The  hourly  curves  for  nitrogen 
and  phosphorus  elimination  during  the  feeding  of  brain  were 
much  the  same. 

Kolpakcha  (1888)  studied  the  source  of  the  nitrogen  of  the 
urine  in  8  extensive  balance  experiments  with  dogs,  by  comparison 
.of  the  relative  amounts  of  the  various  constituents  of  the  food  with 
the  relative  amounts  of  these  same  constituents  in  the  tissues  of 
the  animal,  and  in  the  excreta.  The  author  gives  formulae  for  de- 
termining mathematically  the  proportion  of  food-,  circulating-  and 
tissue-protein  katabolized. 


PHOSPHOEUS  METABOLISM  217 

Among  the  conclusions  reached  by  the  author  are  the  follow- 
ing: In  fasting,  two  kinds  of  protein  are  broken  down,  (1)  "stored 
protein,"  or  protein  stored  in  the  body,  after  excessive  feeding,  that 
has  not  had  time  to  become  part  of  the  tissue  protein,  and  (2)  tissue 
protein,  or  protein  that  is  really  a  part  of  the  tissue  of  the  body. 
The  former  is  said  to  be  broken  down  with  comparative  ease,  the 
latter  being  rather  stable.  From  the  ratio  of  phosphoric  acid  to 
nitrogen  in  the  urine  it  is  considered  that  the  cleavage  of  tissue  pro- 
tein begins  on  the  first  day  of  fasting,  though  it  is  stored  protein 
especially  which  is  broken  down.  As  the  stored  protein  is  exhaust- 
ed the  organism  approaches  a  condition  in  which  it  must  live  entire- 
ly on  its  own  tissues.  The  author  believes  that  the  increased 
amount  of  phosphoric  acid  observed  in  the  urine  after  a  fast  of  con- 
siderable duration,  or  when  the  animal  is  receiving  an  insufficient 
amount  of  food,  is  due  to  autolysis  of  the  bones,  since  there  is  dur- 
ing these  conditions  an  increase  in  the  proportion  of  alkaline  earth 
phosphates  to  alkali  phosphates  in  the  urine.  He  also  believes,  in 
harmony  with  Forster's  conclusions,  that  when  the  organism  is  sup- 
plied with  food  poor  in  phosphoric  acid  it  retains  some  of  the  phos- 
phoric acid  from  the  protein  which  is  broken  up. 

Von  Tschirwinski  (1889)  studied  the  relative  development  of 
the  bones  and  teeth  of  sheep,  and  found  results  not  in  harmony  with 
the  theory  of  an  interdependence  between  the  time  of  cutting  teeth 
and  the  development  of  the  bones. 

Bergell  (1898b)  states  that  normal  storage  of  nitrogen  and 
phosphorus  is  marked  by  a  relation  of  one  to  the  other  like  that  in 
muscle. 

Von  Moraczewsky  (1901)  showed  that,  in  a  6-day  balance  peri- 
od, with  a  case  of  acromegaly,  the  phosphorus  retention  was  double 
the  amount  of  the  calcium  retention.  A  6-day  period,  however, 
is  insufficient  for  reliable  conclusions  as  to  mineral  metabolism. 

Siven  (1901)  notes  the  lack  of  parallelism  between  nitrogen  and 
phosphorus  metabolism,  which  fact  he  relates  to  the  difference  of 
origin  of  these  constituents,  since  the  outgoing  phosphorus  is  de- 
rived in  part  each  from  organic  and  inorganic  phosphorus  com- 
pounds in  the  body. 

Dapper  (1902)  investigated  the  question  as  to  the  method  of 
retention  of  nitrogen  in  a  man  26  years  old.  Data  from  this  work 
are  below.  In  periods  II  and  III  the  ratios  of  N:P205  in  the 
amounts  retained  were  1.69:1  and  1.24  :1,  respectively,  while  the 
ratio  in  muscles  is  7.6 :1.  The  considerable  amounts  of  calcium  and 
phosphorus  retained  must  have  been  deposited  largely  in  the  bones. 


218 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


AVERAGE    DAILY    NITROGEN,    CALCIUM    AND    PHOSPHORUS    BAL- 
ANCES  WITH  A  MAN  TWENTY-SIX  YEARS  OLD— Grams 


Intake 

Excreta 

Balances 

Periods 

N 

P2O5 

CaO 

N 

P2O5 

CaO 

N 

P2O5 

CaO 

I 
6  days.... 

II 
12  days.... 

Ill 
9  days.... 

20.247 
20.093 
24.581 

5.111 
7.033 
7.806 

3.547 
4.818 
4.891 

18.07 

16.774 

22.093 

5.617 
5.022 
5.746 

4.237 
3.465 
4.375 

+2.18 
+3.32 
+2.55 

—0.409 
+1.961 
+2.059 

-0.690 
+1.354 
+0.495 

In  balance  experiments  by  Tangl  (1902a,  1902b)  with  horses 
(see  Phos.  Req.  of  Horses)  we  see  a  case  of  deficiency  of  phosphorus 
in  a  ration  limiting  calcium  retention,  the  calcium  not  being  retained 
in  proportion  to  absorption.  On  the  hay  ration  the  phosphorus 
was  insufficient,  while  there  was  considerable  calcium  retention. 
In  the  oats  and  hay  ration,  with  a  reduced  calcium  intake,  there  was 
an  increased  calcium  storage,  because  of  the  presence  in  this  ration 
of  phosphorus  sufficient  to  cause  retention  and  allow  of  the  deposi- 
tion of  calcium  phosphate  in  the  bones. 

Gilbert  and  Posternak  (1903)  submitted  figures  showing  that 
the  proportion  of  nitrogen  to  phosphorus  in  the  urine  is  so  variable 
that  we  must  consider  the  phosphorus  requirement  as  in  large  meas- 
ure independent  of  that  for  nitrogen. 

Ehrstrom  (1903a),  (see  Nutr.  Val.  Org.  and  Inorg.  P.)  from 
balance  experiments  on  a  man,  concluded  that  phosphorus  and  ni- 
trogen metabolism  need  not  run  parallel,  and  also  that  the  organism 
has  not  the  same  tendency  to  establish  a  phosphorus  equilibrium 
that  it  has  a  nitrogen  equilibrium,  regarding  which  conclusions 
there  is  no  doubt,  since  they  simply  signify  that  the  organism  can 
store  phosphorus  in  the  bones  in  large  measure  irrespective  of  the 
status  of  the  nitrogen  metabolism. 

Liithje  and  Berger  (1904)  reported  results  of  five  balance  ex- 
periments with  human  beings  having  for  their  object  the  study  of 
nitrogen,  phosphorus  and  calcium  metabolism  in  convalescents  from 
typhus  fever.    Data  from  this  work  are  on  the  following  page. 

In  these  experiments  the  object  was  to  bring  about  maximum 
nitrogen  retention,  and  then,  by  a  comparison  of  the  constituents  of 
the  food  and  excreta,  to  determine  the  form  in  which  the  nitrogen 
had  been  stored.  The  authors'  computations  were  based  on  Katz's 
analyses  of  muscle  and  Zalesky's  analyses  of  bone. 


PHOSPHOKUS  METABOLISM 


219 


AVERAGE  DAILY  RETENTION  OF  NITROGEN,  CALCIUM  AND  PHOS- 
PHORUS  BY  MATURE  MEN  ON  HEAVY  FEEDING— Grams 


Condition  of  subject 

Diet 

N 
Intake 

P2O5 
Intake 

CaO 

Intake 

N 
Bal- 
ance 

P2O5 
Bal- 
ance 

CaO 
Bal- 
ance 

Weight 

of 
subject 

Length 
of  period 
in  days 

Typhus  convalescent, 
age  30  years, 
reduced  state 

Milk,  nutrose, 
sugar 

40.246 

10.887 

5.740 

+10.999 

+2.832 

+1.702 

46.9 
49.6 

10 

Typhus  convalescent, 
age  24  years, 
well  nourished 

Milk,  nutrose, 
sugar,  zwei- 

back 

28.337 

8.052 

5.628 

+5.165 

+1.291 

+0.741 

63.3 
66.3 

12 

Typhus  convalescent, 

same  as  in  first  period, 

16  days  later 

Milk,  sugar, 
nutrose 

54.583 

12.385 

7.760 

+11.419 

+1.255 

+0.454 

57.2 
58.1 

7 

Normal 

Milk,  sugar, 
nutrose 

27.337 

9.305 

5.339 

+4.566 

+3.381 

+1.637 

77.5 
77.0 

7 

Normal;   same  as  in 

first  and  third  periods, 

after  recovery 

Milk,  sugar, 
nutrose 

57.187 

12.497 

5.910 

+6.768 

+1.747 

+0.144 

59.2 
61.7 

10 

From  the  above  data  Liithje  and  Berger  compute  that  in 
periods  1  and  2  nitrogen,  calcium  and  phosphorus  were  retained  in 
proportion  for  the  formation  of  flesh  and  bone ;  that  in  period  3  there 
was  nitrogen  retention  without  the  corresponding  quantity  of  phos- 
phorus ;  and  that  in  periods  4  and  5  there  was  phosphorus  retention 
in  excess  of  that  which  could  be  used  with  the  other  food  constitu- 
ents in  the  formation  of  flesh  and  bone. 

They  conclude,  therefore,  that  with  a  large  retention  of  nitro- 
gen in  a  relatively  short  time,  there  is  usually  a  quantity  of  phos- 
phorus retained  which  corresponds  to  the  nitrogen  as  does  the  phos- 
phorus of  flesh  to  the  nitrogen  of  flesh ;  but  there  can  also  be  an  ex- 
cess of  nitrogen  retained,  and  this  is  utilized  in  a  different  way,  per- 
haps in  the  form  of  albumin,  as  a  dead  cell-inclusion,  corresponding 
to  the  cell-deposit  of  glycogen  and  fat;  and  finally,  in  other  cases, 
more  phosphorus  may  be  retained  than  that  corresponding  to  the 
relation  of  nitrogen  to  phosphorus  in  flesh. 

L.  F.  Meyer  (1904a,  1904b)  found,  in  balance  experiments  with 
dogs,  that  nitrogen  could  be  retained  from  proteins  which  were  so 
poor  in  phosphorus  as  to  be  incapable  of  maintaining  phosphorus 
equilibrium.  His  data  show,  however,  that  the  nitrogen  retention 
was  greater  when  the  phosphorus  balance  was  positive  than  when  it 
was  negative,  the  nitrogen  intake  remaining  more  nearly  constant 
than  the  phosphorus.  Meyer  also  notes  the  inclination  of  the  organ- 
ism to  store  phosphorus  ingested  in  excess  of  the  maintenance  re- 
quirement, rather  than  to  maintain  an  equilibrium,  as  is  the  case 
with  regard  to  nitrogen. 


220  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Gumpert  (1905)  (see  Sanatogen,  feeding  exp.)  shows,  in  peri- 
ods of  3  and  4  days,  a  retention  of  P205  of  0.143  and  0.600  gm.  co- 
incident with  a  loss  of  CaO  of  0.307  and  0.244  gm. 

Trowbridge  and  Woodman  (1909)  found  that  young,  growing 
steers  continued  to  grow  in  height,  and  to  build  up  skeleton  even 
when  losing  in  weight,  but  the  extent  to  which  this  involved  redis- 
tribution of  body  constituents  was  not  shown. 

A  large  measure  of  independence  of  growth  of  skeleton  and  soft 
parts  has  been  demonstrated  by  Fleischner  (1906)  with  infants ;  by 
Waters  (1908, 1909, 1909-10)  with  cattle,  and  by  Aron  (1910b)  with 
dogs. 

Kinberg  (1911),  in  a  study  of  nitrogen  hunger,  considered  the 
proportions  of  nitrogen,  sulphur  and  phosphorus  in  the  urine.  These 
relations  varied  irregularly  throughout  the  period,  but  within  com- 
paratively narrow  limits,  viz.,  N:H2S04  as  5.4 — 5.6:1  and  N:P205  as 
4.6—6.6:1. 

Lipschiitz  (1911a)  and  many  others  express  the  opinion  that  in 
case  of  lack  of  a  required  element  in  the  food,  the  body  takes  from 
less  important  parts  to  supply  the  more  important.  An  important 
observation  of  recent  date,  in  this  field,  is  that  of  Gregersen  (1911) 
who  determined  in  metabolism  experiments  with  rats  that  even 
with  an  abundant  intake  of  phosphorus  in  assimilable  form  no  phos- 
phorus is  retained  from  a  nitrogen-free  diet,  thus  suggesting  a  com- 
plete final  dependence  of  the  metabolism  of  phosphorus  upon  that  of 
nitrogen. 

Summary.  From  a  consideration  of  the  above  notes  we  are 
able  to  understand  that  nitrogen  and  sulphur  are  somewhat  closely 
interdependent,  since  they  occur  associated  in  the  same  compounds ; 
phosphorus  is  to  a  considerable  extent  independent  of  nitrogen  be- 
cause of  its  relative  abundance  in  the  skeleton,  while  calcium  is  still 
more  independent  of  nitrogen  because  of  the  poverty  of  the  nitrog- 
enous soft  parts  in  calcium.  Phosphorus  is  to  some  extent  inde- 
pendent of  calcium  because  of  its  association  with  nitrogen  in  the 
soft  parts,  but  calcium  is  closely  dependent  on  phosphorus  because 
it  is  not  stored  in  considerable  amounts  except  as  combined  with 
phosphorus. 

We  must  regard  with  considerable  skepticism  attempts  to  de- 
termine with  much  particularity  the  sources  of  excreted  constitu- 
ents by  their  relative  amounts. 


PHOSPHORUS  METABOLISM  221 

METABOLISM  EXPERIMENTS  WITH  INORGANIC 
PHOSPHATES 

J.  Lehmann  (1859)  found  in  balance  experiments  with  a  calf 
that  alkaline  earth  phosphates  could  be  absorbed  and  retained.  He 
concluded  that  many  foods  were  lacking  in  bone-forming  constitu- 
ents, and  that  hay  was  the  best  bone-forming  food. 

Von  Gohren  (1861)  conducted,  a  balance  experiment  with  a 
lamb  on  a  ration  of  meadow  hay,  to  which  was  added,  in  one  period, 
calcium  and  magnesium  phosphates.  Data  from  this  test  are  be- 
low. 

AVERAGE  DAILY  BALANCE  DATA  FROM  A  LAMB  ON  A  RATION  OF 

MEADOW  HAY  WITH  AND  WITHOUT  ADDED   CALCIUM  AND 

MAGNESIUM  PHOSPHATES— Grams 


Period 
and  days 

Ration 

Food 
CaO 

Food 
MgO 

Food 
P2O5 

Balance 
CaO 

Balance 
MgO 

Balance 
P2O5 

I — 7  days.. 
11— 6  days.. 

Meadow  hay 

Meadow  hay,  10  g-m.  alkaline 
earth  phosphates 

7.081 
10.421 

2.395 
2.428 

3.187 
7.440 

+0.053 
+1.183 

+0.164 
+0.275 

+0.227 
+1.821 

These  data  show  that  lambs  will  digest  and  retain  very  much 
more  calcium,  magnesium  and  phosphorus  than  was  present  in  this 
meadow  hay,  if  these  elements  be  added  to  the  hay  as  alkaline  earth 
phosphates. 

Two  lots  of  lambs,  of  four  each,  were  fed  on  these  same  rations 
for  45  days.  The  lot  which  received  the  phosphates  gained  in 
live  weight  7.2  percent  more  than  the  one  which  received  no  phos- 
phates. VonGohren  noted  that  the  urine  of  lambs  is  practically 
free  from  phosphorus  and  that  the  calcium  also  is  found  mostly  in 
the  feces. 

Blondlot  (1861)  states  that  the  administration  of  doses  of  0.125 
gm.  sodium  hypophosphite  produces  no  phosphorous  acid  test  in  the 
urine,  but  that  doses  of  0.50  gm.  give  the  characteristic  green  flame. 

Gamgee,  Priestley  and  Larmuth  (1876-77)  compared  the  so- 
dium salts  of  pyro-,  ortho-  and  meta-phosphoric  acids  by  subcutan- 
eous, intravenous  and  oral  administration  to  frogs,  rabbits  and  dogs. 
The  ortho-  salt  was  found  to  be  inert,  the  pyro-  salt  markedly  poison- 
ous to  the  heart,  when  introduced  into  the  circulation,  and  the  meta- 
salt  also  poisonous  but  to  a  less  marked  degree. 

Paquelin  and  Joly  (1877)  administered  2  gm.  of  sodium  pyro- 
phosphate daily  for  5  days  to  a  woman,  and  found  during  this  peri- 
od and  the  following  5  days  that  apparently  all  of  the  pyrophospho- 


222 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


ric  acid  was  eliminated  in  the  urine  unchanged.  The  sodium  pyro- 
phosphate had  a  slight  diuretic  effect.  One  qualifying  circum- 
stance, however,  requires  mention.  The  diet  was  not  maintained 
entirely  uniform  during  this  experiment. 

Paquelin  and  Joly  (1878)  administered  to  a  woman  during  5 
days  a  total  of  5  gm.  of  sodium  hypophosphite.  They  concluded 
that  the  hypophosphites  also  are  excreted  unchanged  in  the  urine, 
and  that  they  have  a  diuretic  effect. 

Vermeulen  (1884)  studied  the  physiological  action  of  hypophos- 
phites, but  without  important  results.  He  believes  that  they  pass 
through  the  body  unchanged.  He  determined  hypophosphites  in 
urine  by  first  removing  the  phosphates  by  uranium  precipitation  and 
then  determining  total  phosphates  in  a  KC103+HC1  digest  of  the 
filtrate. 

J.  Neumann  (1893b)  published  results  of  two  calf-feeding  ex- 
periments, in  one  of  which  calcium  phosphate,  and  the  other  cal- 
cium carbonate,  was  added  to  a  skim  milk  diet.  The  calf  used  in  the 
first  experiment  was  5*/2  weeks  old,  and  its  weight  at  the  beginning 
of  the  experiment  was  65.63  kg.  The  calf  used  in  the  second  trial 
was  8  weeks  old,  and  weighed  80.87  kg.  at  the  beginning. 

AVERAGE    DAILY    CALCIUM    AND  PHOSPHORUS  BALANCES    WITH 

CALVES  ON  A  DIET  OF  SKIM  MILK  AND  MINERAL 

SUPPLEMENTS— Grams 


Date  of 
periods 

CaO 

Intake 

CaO 

Outgo 

CaO 
Balance 

P2O5 
Intake 

P2O5 
Outgo 

P2O5 
Balance 

Diet 

Sept.  10-14.. 
Sept.  15-16.. 
Sept.  17-19.. 

24.63 
27.38 
29.19 

12.86 
14.42 
16.77 

+11.77 
+12.96 
+12.42 

30.46 
32.83 
34.33 

14.91 
16.87 
18.39 

+15.55 
+15.96 
+15.94 

15000  gm.  skim  milk. 

Same  plus  7.5  gm.  calcium  phosphate. 

Same  plus  12  g-m.  calcium  phosphate. 

Sept.  29-30.. 
Oct.  1-2.... 
Oct.  3-7.... 
Oct.  8-9.... 
Oct.  10-14.. 

26.252 

27.478 
30.008 
26.407 
26.412 

14.562 

14.854 
15.820 
15.285 
14.776 

+11.690 
+12.624 
+14.188 
+11.122 
+11.636 

34.039 
33.863 
33.980 
34.240 
32.247 

19.511 
19.712 
19.327 
19.696 
19.947 

+14.528 
+14.151 
+14.653 
+14.544 
+14.300 

16000  gm.  skim  milk. 

16000  gm.  skim  milk. 

Same  plus  7.5  gm.  calcium  carbonate. 

16000  gm.  skim  milk. 

16000  gm.  skim  milk. 

Thus  it  appears  that  both  calcium  carbonate  and  phosphate, 
when  added  to  skim  milk,  are  digested  and,  in  considerable  meas- 
ure, retained  by  the  young  calf ;  and  further  that  calcium  carbonate 
increased,  not  only  the  calcium  storage,  but  also  the  phosphorus  re- 
tention. Neither  the  calcium  nor  the  phosphorus  of  the  phosphate 
were  so  well  retained  as  the  same  elements  in  the  milk,  but  the  cal- 
cium of  the  carbonate  was  retained  in  about  the  same  proportion  as 
the  calcium  of  the  milk.  It  was  found  that  both  of  these  mineral 
supplements  tended  to  reduce  the  gain  in  live  weight. 


PHOSPHORUS  METABOLISM  223 

Weiske  (1895b)  mentions  experiments  by  Graff  enberger  show- 
ing that  on  an  oat  ration  rabbits  digested  a  little  more  protein,  fat, 
crude  fiber  and  nitrogen-free  extract  without  tricalcic  phosphate 
than  when  this  salt  was  added  to  the  diet ;  and  also  experiments  of 
his  own  where  calcium  carbonate  was  added  to  a  ration  of  meadow 
hay,  with  rabbits,  in  which  the  mineral  supplement  seems  to  have 
reduced  to  a  slight  extent  (1.35  percent)  the  digestibility  of  the  pro- 
tein, but  to  have  increased  to  the  extent  of  10.11  percent  the  digesti- 
bility of  the  nitrogen-free  extract. 

Boddaert  (1896)  found  calcium  and  sodium  hypophosphites 
rapidly  excreted,  apparently  as  such,  in  the  urine,  whether  adminis- 
tered per  os  or  subcutaneously,  with  rabbits,  dogs  and  men. 

Massol  and  Gamel  (1901)  added  sodium  phosphate  to  a  solution 
of  calcium  hypophosphite.  The  solution  was  then  made  alkaline, 
and  a  precipitate  of  tricalcium  phosphate  was  formed,  leaving  so- 
dium hypophosphite  in  solution.  The  same  reaction,  they  state, 
takes  place  in  the  intestine,  and  the  tricalcic  phosphate  is  lost  to  the 
organism.  The  sodium  hypophosphite  is  absorbed,  and  excreted 
unchanged  by  the  kidneys,  there  being  no  oxidation  of  phosphorous 
acid.  Thus  calcium  hypophosphite  removes  phosphorus  from  the 
system  in  proportion  to  the  amount  fed. 

Panzer  (1902),  in  studying  the  fate  of  calcium  hypophosphite 
in  the  body,  reached  conclusions  by  the  feeding  of  the  compound, 
and  then  by  qualitative  tests  (the  green  flame  with  nascent  hydro- 
gen, and  the  brown  silver  phosphide  precipitate)  in  the  urine,  feces, 
blood  and  organs.  The  tests  were  made  in  part  on  a  man  and  in 
part  on  dogs. 

Panzer  decided  that  if  the  methods  were  adequate  to  this  in- 
vestigation calcium  hypophosphite  fed  to  a  dog  is  quickly  and  almost 
completely  absorbed,  passes  through  the  organism  without  being 
held  back  anywhere,  and  is  completely  eliminated  within  24  hours. 

Martinet  (1902a)  says  that  hyperacidity  of  the  urine  accom- 
panied by  abnormally  small  amount  of  phosphates  is  frequently  an 
index  of  hypochlorhydria  in  the  stomach,  and  that  in  such  cases 
the  administration  of  phosphoric  acid  may  promote  digestion  and 
check  fermentation  by  its  eupeptic  and  antiseptic  action. 

In  a  later  article  (1905)  Martinet  writes  that  by  the  use  of  phos- 
phoric acid  in  48  cases  of  dyspepsia,  32  cures  have  been  accom- 
plished, the  liver  also  being  benefited ;  urea  excretion  is  increased ; 
indican  disappears  and  glycosuria  of  hepatic  origin  is  relieved ;  also 
the  hepato-pancreatic  function  is  stimulated;  intestinal  functions 
are  regulated,  and  diarrhoea  suppressed.  Treatment  with  sodium 
carbonate  inhibits  the  digestive  functions,  especially  of  the  stomach, 
while  the  phosphoric  acid  treatment,  on  the  contrary,  stimulates 
these  functions. 


224  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Gouin  and  Andouard  (1902-3)  conducted  digestion  experiments 
on  a  heifer,  having  for  their  object  a  study  of  the  effects  on  metabo- 
lism of  certain  inorganic  salts.  The  basal  ration  was  composed  of 
milk,  rice  meal,  clover,  peas,  and  oats.  Acid  potassium  phosphate,  in 
the  proportion  of  5  grams  per  100  kg.  live  weight,  was  fed  during  8 
weeks.  Benefit  was  thought  to  have  resulted.  The  feeding  of 
150  grams  of  powdered  bone  per  day,  equivalent  to  28.29  gm.  P205, 
greatly  increased  the  urinary  phosphorus,  and  had  a  depressing  ef- 
fect on  the  digestion  of  protein,  carbohydrates  and  fat. 

Cautru  (1903, 1904a,  1904b)  fed  pure  phosphoric  acid  in  consid- 
erable quantities  to  guinea  pigs,  dogs,  ducks,  horses  and  human  be- 
ings, during  protracted  periods,  to  determine  limits  of  toleration  and 
therapeutic  effects.  In  large  doses,  even,  the  acid  is  innocuous,  and 
is  recommended  by  Cautru  for  use  in  such  morbid  states  as  are  char- 
acterized by  demineralization  and  hypoacidity  of  the  urine.  As 
much  as  2-4  gm.  daily  were  fed  to  human  beings  for  more  than  5 
years ;  a  dog  weighing  8.5  kg.  was  given  gradually  increasing  doses 
up  to  3  gm.  for  one  month;  a  duck  was  given  one  gram  per  day; 
horses  were  given  30-100  gm.  daily;  two  guinea  pigs  weighing  350 
gm.  received  0.5-1.0  gm.  daily,  etc. 

Kohler  et  al.  (1904),  in  experiments  with  yearling  lambs,  found 
that  when  added  to  a  ration  containing  comparatively  little  mineral 
matter  tricalcic  phosphate  was  retained  in  larger  proportion  to  the 
intake  than  were  dicalcic  phosphate,  precipitated  bone  earths  (di- 
and  tri-salts),  or  calcined  bone.  After  making  this  observation,  he 
determined  with  a  6-months-old  lamb  that  the  reason  the  dicalcic 
phosphate  was  not  so  well  retained  as  the  tricalcic  salt  was  its  lack 
of  calcium.  When  additional  calcium  was  provided,  as  calcium  lac- 
tate, the  phosphorus  of  the  dicalcic  salt  was  much  more  efficiently 
retained  than  that  of  the  tricalcic  phosphate. 

Of  the  phosphorus  of  the  precipitated  bone  earths  the  retention 
was  13.1  percent,  of  the  calcined  bones  14.2  percent,  of  the  dicalcic 
phosphate  26  percent  and  of  the  tricalcic  salt  35.5  percent.  With 
the  younger  lamb  the  phosphorus  of  the  tricalcic  phosphate  was  re- 
tained to  the  extent  of  38.6  percent  of  the  intake,  of  the  dicalcic 
phosphate,  35  percent  when  fed  alone,  and  54.7  percent  when  fed 
with  calcium  lactate. 

Joulie  (1904)  administered  phosphoric  acid  in  rheumatism  of 
horses.  The  dose  was  10  c.c.  phosphoric  acid  (sp.  gr.  1.35) 
diluted  to  a  liter.  He  states  that  such  introduction  of  phosphoric 
acid  diminishes  the  alkalinity  of  the  blood,  and  increases  the  elim- 
ination of  calcium  phosphate  by  the  kidneys. 


PHOSPHORUS  METABOLISM 


225 


Kramer  (1907)  investigated  the  effects  of  ingestion  of  calcium 
phosphate  on  the  laxative  tendency  of  beet  leaves  when  fed  to  cows. 
It  is  usually  credited  with  preventing  a  laxative  effect,  but  in  this 
case  the  beet  leaves  were  not  sufficiently  laxative  to  permit  of  this 
observation. 

Kohler,  Honcamp  and  Eisenkolbe  (1907)  reported  results  on 
further  tests  of  inorganic  phosphates  with  lambs.  The  authors 
conclude  that  the  phosphorus  of  the  dicalcic  salt  is  slightly  better 
absorbed  and  retained  than  the  tricalcic  salt,  and  that  the  addition 
of  calcium  lactate  or  calcium  carbonate  is  without  influence  on  the 
degree  of  retention  of  the  phosphorus.  The  results  of  these  tests 
are  so  variable  that  it  seems  to  the  writers  unwise  to  draw  conclu- 
sions from  them. 

W.  Muller  and  vonWendt  (1908)  found  that  precipitated  cal- 
cium phosphate  fed  to  cattle  suffering  with  diarrhoea,  from  eating 
partially  decomposed  beet  leaves,  served  to  allay  the  symptoms, 
while  calcium  carbonate  accentuated  the  difficulty.  The  authors 
explain  the  unfavorable  effect  of  the  carbonate  as  due  to  the  neutral- 
ization of  the  acid  products  of  bacterial  growth,  thus  favoring  fur- 
ther growth.  Oxalic  acid  was  shown  not  to  be  the  cause  of  the  in- 
digestion. 

Aron  and  Sebauer  (1908)  concluded  from  feeding  experiments 
with  rabbits  and  dogs,  using  calcium-poor  rations,  to  which  in  some 
cases  bone  meal,  calcium  phosphate  or  calcium  carbonate  was  added, 
that  the  lack  of  these  calcium  compounds  affected,  in  noteworthy  de- 
gree, only  the  bones,  these  being  below  normal  in  dry  substance  and 
in  the  calcium  content  of  the  dry  matter,  the  reduction  of  calcium 
content  of  the  ash  being  considered  immaterial.  The  calcium  con- 
tent of  the  flesh  and  blood  shows  no  reduction,  while  the  calcium 
content  of  the  brain  is  reduced  but  little. 

Dibbelt  (1909)  conducted  metabolism  experiments  with  dogs, 
in  which  dicalcic  phosphate,  sodium  chloride  and  magnesium  sul- 
phate were  added,  in  different  periods,  to  a  standard  basal  ration.  A 
part  of  the  data  are  below. 

AVERAGE   DAILY   CALCIUM    AND    PHOSPHORUS    BALANCES    WITH 
DOGS  AS  AFFECTED  BY  INORGANIC  SALTS— Grams 


Exp. 
No. 

Length 
of  period 
in  days 

Gain  in 

weight 

CaO 

Intake 

CaO 

Retention 

P2O5 
Intake 

P2O5 

Retention 

Diet  and  treatment 

1 
2 

3 

4 

15 
10 

9 

11 

100 

80 

0.0500 
0.0500 

0.0500 
0.2178 

-0.0478 
+0.010 

—0.001 
-1-0.1388 

2.3124 
2.3124 

2.3124 

2.5188 

0.6181 

1.1882 

0.7384 
1.0989 

Horse  meat  and  pork  fat. 

Horse  meat  and  pork  fat;  immediately 

following-  Exp.  No.  1. 
Horse  meat,  pork  fat  and  12  gm.  salt. 
Horse  meat,  pork  fat  and   0.5   g-m. 

dicalcium  phosphate. 

5 
6 

7 
23 

3.57 
9.78 

0.0150 
0.0160 

+0.003 
+0.0062 

0.6937 
0.7400 

0.1907 
0.3618 

Horse  meat,  fat  and  salt. 
Horse  meat,  fat,  salt  and  magnesium 
sulphate. 

226  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

In  Exp.  No.  1  it  appeared  that  horse  meat  and  pork  fat  were 
deficient  in  calcium,  but  in  Exp.  No.  2,  with  the  same  diet  and  same 
animal,  it  seemed  that  the  dog  had  accomplished  an  adjustment  to 
this  diet.  The  result  in  No.  3,  where  salt  was  fed,  was  essentially 
the  same,  so  far  as  calcium  is  concerned;  at  least  it  did  not  differ 
from  No.  2  as  much  as  No.  2  differed  from  No.  1.  In  Exp.  No.  4, 
however,  there  was  a  marked  calcium  storage  coincident  with  the 
feeding  of  dicalcium  phosphate. 

Experiments  5  and  6  were  on  another  dog.  There  appeared 
to  be  an  effect  by  the  magnesium  sulphate  to  increase  the  storage 
of  calcium  and  phosphorus,  but  the  evidence  must  be  regarded  as 
inconclusive  on  this  point.  The  preliminary  period  was  short. 
Some  days  after  the  conclusion  of  Exp.  6  this  dog  was  killed,  and 
the  calcium  content  of  its  tissues  was  compared  with  the  same  from 
another  dog  which  had  received  the  same  ration  except  that  dical- 
cium phosphate  replaced  magnesium  sulphate.  The  soft  parts  of 
the  dog  receiving  magnesium  sulphate  were  found  much  richer,  but 
the  bones  poorer,  in  calcium,  than  those  of  the  dog  which  received 
dicalcium  phosphate.  These  differences  the  author  explains  by 
assuming  that  the  magnesium  sulphate  had  reacted  with  the  sol- 
uble calcium  phosphates  to  form  the  comparatively  insoluble  cal- 
cium sulphate. 

Dibbelt  conducted  other  experiments  in  the  artificial  production 
of  rachitic  conditions,  and  concluded  that  the  principal  difference 
between  these  conditions  and  rachitis  is  in  the  cause  of  the  calcium 
deficiency,  this  being  due  in  the  latter  case  especially  to  anomalies  of 
absorption  and  secretion,  rather  than  to  calcium  shortage  in  the 
food  as  a  first  cause. 

Crawford  (1910)  fed  sodium  metaphosphate,  sodium  pyrophos- 
phate and  sodium  orthophosphate  to  rabbits  in  a  study  of  cotton- 
seed meal  poisoning.  The  feeding  of  2  gm.  of  the  metaphosphate 
daily  for  two  days  caused  the  death  of  the  subject,  as  also  did  the 
feeding  of  0.5  gm.  of  the  pyrophosphate  per  day  for  7  days.  The  dos- 
age was  enormous.  The  author's  conclusion  that  cottonseed  meal  poi- 
soning was  due  to  pyrophosphates  has  not  withstood  criticism. 

Delaini  (1911)  submits  data  showing  that  human  beings  and 
dogs  eliminate  in  the  urine  practically  the  whole  amount  of  hypo- 
phosphite  of  sodium  administered  either  per  os  or  subcutaneously, 
but  that  rabbits  otherwise  dispose  of  perhaps  20  percent  of  the  in- 
take. 

Patta  (1910)  administered  to  a  dog,  by  injection,  varying  doses 
of  sodium  hypophosphite.  If  administered  in  large  amounts  about 
half  of  all  introduced  can  be  recovered,  unoxidized,  in  the  urine; 


PHOSPHORUS  METABOLISM  227 

when  smaller  amounts  are  administered,  the  proportion  recovered  is 
less,  being  as  low  as  one-sixth  at  times.  Hence  it  is  maintained 
that  the  phosphorus  of  hypophosphites  can  be  retained  by  the  or- 
ganism. 

Starkenstein  (1911)  theorizes  to  the  effect  that  the  reason  that 
pyrophosphoric  acid  and  metaphosphoric  acid  are  poisonous  when 
injected  into  the  organism,  while  orthophosphoric  acid  is  not,  is 
that  these  acids  are  formed  from  orthophosphoric  acid  by  condensa- 
tion of  molecules,  and  loss  of  water.  Experiment  showed  that  if 
orthophosphoric  acid  be  used  in  sufficient  amount  and  concentra- 
tion, it  also  poisons — 10  c.c.  of  14  percent  Na,HP04,  injected,  pro- 
duces the  same  effect  as  2  c.c.  of  5  percent  pyrophosphate  solution. 
The  poisonous  salts  are  more  alkaline.  Is  the  poison  due  to  alka- 
linity? Blake  found  that  they  are  more  easily  dissociated,  which 
would  mean  greater  alkalinity.  Dissociation  tests  of  the  acids  by 
reaction  toward  different  indicators,  by  conductivity,  and  by  heat  of 
reaction  all  show  that  the  pyrophosphoric  acid  has  higher  true  acidi- 
ty than  the  orthophosphoric  acid.  Two  of  the  H+  of  the  pyrophos- 
phoric acid  split  off  as  easily  as  one  of  the  orthophosphoric  acid; 
therefore  probably  the  tetrapyrophosphate  of  sodium  dissociates  to 
give  as  much  OH"  as  does  the  tertiary  salt  of  the  ortho  acid. 

Withers  and  Ray  (1913)  submit  evidence  controverting  the 
idea  that  cottonseed  meal  poisoning  is  due  to  pyrophosphates. 

See  also  Etzinger  (1874)  on  digestibility  of  bones  by  dogs; 
Charrin  and  Desgrez  (1896)  on  subcutaneous  injection  of  salts,  in- 
cluding sodium  phosphate;  Vosgien  and  Geroline  (1899)  on  reten- 
tion of  inorganic  phosphates  by  rabbits;  Charrin  (1905)  on  subcu- 
taneous injection  of  salts,  including  sodium  phosphate;  Cook  (1909, 
Met.  Phytin),  and  Draeger  (1911)  on  the  assimilation  of  calcium 
phosphates. 

MISCELLANEOUS     EFFECTS     OF     INORGANIC     PHOSPHATES     ON 

GROWTH 

Graffenberger  (1893)  found  that  the  feeding  of  calcium  phos- 
phate to  rabbits  did  not  increase  the  calcium  and  phosphorus  content 
of  their  litters.     , 

Knauthe  (1898)  submits  data  from  experiments  with  carp, 
showing  that  the  addition  of  meat  ash  to  rations  of  meat  meal  and 
corn  meal,  or  meat  meal  and  rice  meal,  serves  to  increase  both  nitro- 
gen retention  and  gain  in  weight. 

W.  P.  Wheeler  (1903)  reported  more  efficient  use  of  food,  and 
more  rapid  growth  of  chicks,  from  the  addition  of  rock  phosphate  to 
a  ration  of  vegetable  foods.  As  a  source  of  calcium  and  phospho- 
rus for  chicks,  however,  Wheeler  recommends  raw  or  cooked  bone. 


228  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Klien  (1908)  found,  in  feeding*  experiments  on  growing  pigs, 
that  when  added  to  a  ration  of  milk,  potatoes,  barley  and  rye,  cal- 
cium carbonate  had  about  the  same  value  as  calcium  phosphate  in 
causing  increase  of  weight. 

Moore,  Roef  and  Knowles  (1908)  found  that  newts  would  re- 
spond to  the  presence  of  both  acid  and  alkali  phosphates  in  the 
water  in  which  they  were  kept,  by  increased  growth,  provided  the 
salts  were  present  in  certain  optimum  concentrations.  In  excess- 
ive amounts  monosodium  phosphate  was  depressant,  the  disodium 
salt  producing  excitation. 

The  same  salts  were  fed  to  guinea  pigs  and  dogs  in  quantities 
sufficient  to  cause  pathological  symptoms,  and  the  histological  con- 
dition of  the  organs  and  tissues  is  described. 

For  other  materials  on  the  effects  of  phosphates  on  general 
growth  see  Mai  (1869),  W.  Cohn  (1870),  J.  Lehmann  (1873,  1877), 
Passon  (1905),  Carlier  (1907,  1909)  and  Forbes  and  associates 
(1914). 

Summary :  Alkaline  earth  phosphates  can  be  absorbed  and  re- 
tained by  sheep  when  added  to  a  ration  of  hay. 

Calcium  phosphate  and  calcium  carbonate  can  be  absorbed  and 
retained  by  the  calf  when  added  to  a  skim  milk  diet.  Calcium  car- 
bonate, under  these  circumstances,  increases  the  phosphorus  reten- 
tion, calcium  thus  appearing  to  be  a  limiting  factor  in  the  use  of 
the  phosphorus  of  milk.  Both  the  carbonate  and  the  phosphate 
tend  to  reduce  the  gain  in  live  weight. 

Tricalcic  phosphate  depressed  to  a  slight  extent  digestibility 
of  oats  by  rabbits,  and  of  a  mixed  ration  of  milk,  grain  and  green 
feeds  by  a  heifer.  Calcium  carbonate  seems  to  depress  the  digest- 
ibility of  protein  and  to  increase  the  digestibility  of  nitrogen-free 
extract  of  oats  with  rabbits. 

Phosphoric  acid  has  been  found  to  stimulate  gastric  functions 
in  cases  of  hypochlorhydric  dyspepsia. 

By  subcutaneous  injection  of  phosphates  it  is  learned  that  the 
ortho-salts,  in  moderate  amounts,  are  inert ;  the  pyro-salts  are  mark- 
edly poisonous;  while  the  meta-salts  are  also  poisonous,  but  to  a 
less  marked  extent. 

Lack  of  consistent  results  makes  difficult  the  use  of  data  on 
the  comparative  values  of  the  various  calcium  salts  fed  to  live  stock. 

We  have  very  little  evidence  that  the  inorganic  salts  of  other 
than  the  ortho-phosphoric  acid  are  of  value  as  bearers  either  of 
phosphorus  or  of  base.  The  hypophosphites  especially  are,  through- 
out therapeutic  literature,  spoken  of  as  inactive. 

In  contradistinction  to  the  effects  of  phosphates  on  the  bones 
the  ingestion  of  inorganic  phosphates  produces  slight  effect  on  body 
growth  in  general. 


PHOSPHORUS  METABOLISM  229 

METABOLISM  OF  NUCLEOPROTEINS  AND  NUCLEIC  ACIDS  * 

GENERAL  DISCUSSION 

Anabolic  Processes.  It  is  evident  from  various  observations 
that  the  animal  organism  is  capable  of  building  up  for  itself  nuclear 
material  from  other  forms  of  protein.  The  first  definite  evidence 
of  such  synthesis  in  the  body  was  that,  from  Miescher's  (1881, 
1896)  observations  on  Rhine  salmon,  where  it  was  concluded  that 
the  nuclein-rich  reproductive  substance  of  these  fish  must  form 
from  the  material  of  the  muscles,  for  nearly  the  entire  development 
of  the  reproductive  organs  and  their  sperm  and  ova  takes  place 
while  the  fish  are  taking  no  food,  the  fibres  of  the  body  muscles  los- 
ing much  of  their  protein  at  the  same  time.  This  view  with  regard  to 
salmon  was  strengthened  by  the  investigations  of  Paton  and  his  as- 
sociates when  they  found  that  the  amounts  of  phosphorus  lost  by 
the  body  muscles  account  quantitatively  for  the  gain  of  phosphorus 
in  the  ovaries  and  testes.  (Paton,  et  ah,  1897-8;  Paton,  1898).  Ac- 
cording to  Milroy  (1908),  it  is  not  true  of  herring  that  the  genitalia 
derive  their  protein  from  the  protein  of  body  muscle,  neither  the 
protein  nor  the  phosphorus  figures  indicating  such  a  relation  in  his 
examinations.  These  studies  were  inconclusive  because  of  the  lack 
of  complete  chemical  accounting  for  the  entire  fish. 

Building  up  nuclein  from  purin-free  protein  must,  according  to 
the  reports  of  Burian  and  Schur  (1897),  take  place  in  the  bodies  of 
suckling  animals.  From  xanthin-base  nitrogen  and  nuclein  phos- 
phorus ,  determinations  on  the  body  substance  of  suckling  pups  at 
different  ages,  and  on  the  mother  dog's  milk,  it  appeared  that  the 
increase  of  nuclein  in  the  young  animal  is  too  great  to  be  accounted 
for  by  the  very  small  amount  of  nuclein  taken  in  the  milk. 

Schmoll  (1904)  discusses  the  chemical  origin  of  leucocytes,  con- 
sidering the  question  of  the  synthesis  of  nucleins  by  the  adult  hu- 
man organism.  Leucocyte  counts  in  the  blood,  and  uric  acid  and 
purin  base  determinations  in  the  urine,  while  patients  were  on  a 
normal  mixed  diet,  or  on  diets  rich  or  very  poor  in  purins,  are  used 
as  evidence  of  the  synthesis  of  leucocytes  (nuclein)  from  the  albu- 
min of  the  food  without  the  necessity  for  food  purins. 

McCollum  (1909)  also  considered  nuclein  synthesis  in  rats.  The 
subjects  were  fed  in  groups  of  3  on  (1)  normal  food,  (2)  a  ration 
free  from  organic  phosphorus  and  from  purins,  (3)  a  ration  such  as 
the  last  with  purins  from  liver  and  hydrolyzed  beef  muscle  added, 
or  (4)  with  casein  added  to  the  basal  ration.      The  bodies  of  some 

*  For  general  discussions  see  A.  Schittenhelm:  Der  NueleinstoftVeehsel ;  Oppenheimer's 
Handbuch,  Vol.  IV,  1st  half,  489-539;  also  Brugsch  and  Schittehhelm  (1910),  and  Schau- 
mann   (1910). 


230  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

were  examined  as  to  percent  of  skeleton  and  percent  of  fat-free  tis- 
sue, but  the  evidence  as  to  the  synthetic  powers  of  the  organism  is 
mainly  the  rate  of  gain  in  live  weight  of  the  animals.  He  con- 
cluded that  the  phosphorus  needed  by  an  animal  for  nuclein  form- 
ation can  be  drawn  from  inorganic  phosphates,  and  that  the  animal 
has  the  power  to  synthesize  the  purin  base  necessary  for  its  nuclein 
formation  from  some  complexes  contained  in  the  protein  molecule, 
and  does  not  necessarily  use  purin  bases  of  exogenous  origin  for 
this  purpose.  Complete  chemical  accountings  for  the  bodies  of 
the  rats  would  have  been  required  to  establish  these  points. 

Other  investigations  involving  the  subject  of  nuclein  synthesis 
are  those  which  are  considered,  in  the  section  on  The  Nutritive  Val- 
ues of  Organic  and  Inorganic  Phosphorus,  as  evidence  of  the  pro- 
duction of  growth  from  rations  which  are  free  from  inorganic  phos- 
phorus. 

Evidence  of  synthetic  changes  in  the  nuclear  material  of  devel- 
oping eggs  of  silk-worms  has  been  submitted  by  Tichomiroff  (1885), 
and  of  hens  by  A.  Kossel  (1885,  1886).  On  the  other  hand,  Shack- 
ell  (1911)  was  unable  to  find  in  the  early  stages  of  developing  sea- 
urchin  eggs  any  convincing  evidence  of  the  synthesis  of  nuclear  ma- 
terial from  the  alcohol-soluble  substances  of  the  cytoplasm ;  and  this 
corresponds  with  the  observations  of  Masing  (1910),  who  com- 
pared unfertilized  sea-urchin  eggs,  or  those  which,  being  fertilized, 
had  just  formed  the  membrane,  with  others  which  had  developed  to 
the  morula  stage,  where  there  are  500-1000  cells.  The  nuclear  mass 
has  at  that  time  multiplied  nearly  100-fold,  but  the  nucleic  acid  con- 
tent was  not  found  to  be  any  greater  than  in  the  unfertilized  eggs. 
Later  (1911b)  he  made  similar  examination  of  the  successive  stages 
of  the  embryos  of  rabbits,  and  of  the  livers  of  embryonal  rabbits. 
Both  the  whole  embryo  and  the  embryo  liver  showed  decreasing 
percentage  content  of  nuclein  as  development  progressed,  though 
the  absolute  amount  increased.  Sections  of  the  livers  showed  under 
the  microscope  a  corresponding  decrease  in  the  relative  size  of  the 
nucleus.  Masing's  work,  then,  makes  it  appear  that  there  is  a 
large  supply  of  nuclear  material  at  the  start  to  provide  for  growth. 

There  is  little  evidence  as  to  the  habit  of  the  body  with  regard 
to  the  details  of  anabolism  during  growth  and  repair.  There  is 
abundant  evidence  that  the  organs  contain  within  themselves  en- 
zymes which  can  break  down  nucleins,  but  whether  they  do  in  fact 
break  them  down  to  the  stage  of  free  purins  and  phosphoric  acid  be- 
fore building  them  into  new  cellular  matter  is  not  known.  It  may 
be  advantageous  and  habitual  for  the  cleavage,  whether  in  the  di- 
gestive tract  or  in  the  tissues,  to  be  stopped  at  an  earlier  stage.  The 


PHOSPHORUS  METABOLISM  231 

investigations  of  Taylor  (1907)  and  of  Robertson  (1907,  1909a, 
1911)  indicate  that  syntheses  may,  under  the  right  circumstances, 
be  brought  about  by  the  same  enzymes  which  under  other  circum- 
stances cause  cleavage. 

The  attempts  which  have  been  made  at  artificial  synthesis  of 
nucleoproteins  or  nucleins  are  reported  and  discussed  in  the  chap- 
ter on  the  chemistry  of  these  compounds. 

Katabolic  Processes.  Three  distinct  methods  of  experimental 
study  have  been  employed  in  attempts  to  learn  of  the  extent  to 
which  nucleoproteins  are  broken  down  in  the  life  processes  of  the 
organism,  the  transformations  involved  and  the  places  at  which 
they  occur. 

First,  isolated  enzymes  or  digestive  secretions  from  the  ali- 
mentary tract  have  been  allowed  to  act  on  the  substances  in  ques- 
tion under  controlled  conditions  and  at  favorable  temperatures,  the 
extent  and  nature  of  the  change  taking  place  in  the  digestive  tract 
of  the  animal  being  judged  by  the  results  of  these  artificial  diges- 
tions, in  vitro.  To  this  may  be  added  the  consideration  of  changes 
which  may  be  brought  about  by  such  bacteria  as  are  found  in  the 
intestine.  It  is  to  be  recognized  that  observations  of  this  kind  lose 
somewhat  in  significance  from  being  made  under  artificial  condi- 
tions ;  but,  on  the  other  hand,  there  is  an  advantage  in  the  limita- 
tion of  variable  or  unknown  factors. 

A  second  type  of  studies  made  outside  the  body,  and  therefore 
under  unnatural  conditions,  but  probably  being  to  an  important  de- 
gree indicative  of  the  body  processes,  has  been  followed  extensively 
in  recent  years.  We  refer  to  studies  with  comminuted  organs  in  sus- 
pension, or  with  water  extracts  of  organs,  as  to  their  autodigestion, 
especially  as  related  to  purins,  and  their  capacities  to  transform  ad- 
ded nucleic  acids  or  their  derivatives.  Evidently  the  organs  and 
tissues  generally  have  within  themselves  enzymes,  or  the  means  for 
producing  them,  by  which  they  may  break  down  and  transform  their 
own  nucleoproteins,  or  partially  digested  nucleoproteins  or  nucleic 
acids  which  may  be  brought  to  them. 

The  third  method  of  study  makes  observations  of  the  actual  re- 
sults in  living  subjects  from  introduction  of  the  substance,  either 
in  the  diet  or  by  some  form  of  injection. 

DIGESTION  STUDIES  WITH  ENZYMES  FROM  THE  ALIMENTARY 

TRACT 

Popoff  (1894)  concluded  from  digestions  of  thymus  gland  with 
ferment  preparations  that '  'Solution  of  nuclein  material  occurs  only 
very  slightly  in  the  stomach,  but  to  a  considerable  extent  in  the  in- 
testine, through  the  agency  of  the  pancreatic  juice.      Here  the  nu- 


232  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

clein  bodies  as  such  are  taken  into  solution,  so  that  it  may  be  in- 
ferred that  they  are  also  resorbed  as  such."  Very  little  inorganic 
phosphorus  appeared  in  the  solution. 

Milroy  (1896)  found  most  of  the  phosphorus  from  natural  or 
artificial  nucleins  in  organic  form  after  trypsin  digestion. 

A.  Neumann's  (1898)  examination  of  the  a-  and  /?-nucleic  acids 
and  nucleothyminic  acid  showed  that  none  of  these  were  attacked 
by  pepsin-hydrochloric  acid,  but  that  they  were  dissolved  by  alka- 
line intestinal  fluids. 

Umber  (1901)  made  long  peptic  and  tryptic  digestions  of  the 
nucleoprotein  from  pancreas.  About  nine-tenths  of  the  nucleopro- 
tein  passed  into  solution  under  the  influence  of  the  pepsin.  The 
pepsin  first  brought  about  a  separation  of  a  protein  from  a  pentose- 
rich  nucleic  acid,  which  was  soluble  and  could  be  absorbed,  but 
which  was  not  further  broken  down  by  pepsin.  The  digestion  of 
the  protein  component,  after  being  split  off  from  the  nucleic  acid, 
proceeds  in  the  ordinary  way  with  the  formation  of  primary  and 
secondary  albumoses,  peptone  and  lower  cleavage  products.  Trypsin 
had  a  like  effect  to  that  of  pepsin,  and  required  less  time. 

Araki  (1903a)  carried  out  artificial  tests  which  seemed  to  indi- 
cate that  the  nuclear  substance  of  the  red  corpuscles  of  bird's  blood 
is  rather  quickly  dissolved  by  trypsin;  that  there  is  in  thymus  ex- 
tract an  enzyme  which  has  a  like  but  slower  action;  also  that  by 
both  trypsin  and  the  thymus  extract  nucleic  acid  is  changed  from  the 
less  soluble  (gelatinizing)  form  which  Kossel  and  Neumann  called 
a-acid  to  the  more  readily  soluble  (non-gelatinizing)  /?-acid,  and 
no  further  digestion  occurs  except  after  long-continued  action ;  and 
that  erepsin  splits  nuclein  and  dissolves  nucleic  acid. 

Nakayama  (1904)  compared  the  action  of  erepsin  from 
a  dog  with  that  of  trypsin  and  of  weakly  alkaline  ex- 
tracts from  the  intestines  of  cattle  and  rabbits.  Commercial 
trypsins  did  not  digest  nucleic  acids.  The  erepsin  slowly  digested 
the  sodium  salts  of  nucleic  acids  from  the  intestine  of  cattle,  from 
thymus,  from  the  spleen  of  cattle,  and  from  the  spermatozoa  of 
Muraenosox  cinereus.  In  each  case  phosphoric  acid  was  found  in 
the  resulting  solution.  The  extract  of  intestine  acted  on  the 
nucleic  acid  much  as  did  the  erepsin. 

In  connection  with  the  study  of  enzyme  action  on  caseinogen 
Plimmer  and  Bayliss  (1906)  tested  the  artificial  digestion  of  yeast 
nuclein  with  trypsin,  and  found  about  four-fifths  of  its  phosphorus 
in  solution  as  phosphoric  acid  after  21  days. 


PHOSPHORUS  METABOLISM  233 

Mitra  (1911)  considered  the  age  at  which  nuclease  appears  in 
the  stomach  of  the  infant.  An  infant  a  year  old  was  fed  on  milk, 
on  milk  with  flour  and  on  porridge  made  from  legumes ;  and  at  inter- 
vals of  0.5, 1, 1.5  and  2  hours  after  -ingestion  portions  were  removed 
from  the  stomach  and  tested  with  connective  tissue  and  muscle  fibre. 
No  nuclease  nor  connectivase  was  found.  A  similar  experiment  on 
a  child  of  fifteen  months  seemed  to  show  the  presence  of  both. 

Levene  and  Medigreceanu  (1911c)  have  made  various  kinds  of 
nucleic  acid  digestion  tests,  among  them  some  such  as  are  now  under 
discussion.  Inosin,  guanosin,  cytidin,  guanylic  acid,  pyrimidin  nucleo- 
tides, yeast  nucleic  acid  and  thymus  nucleic  acid  were  studied.  The 
glucosides  (inosin,  guanosin  and  cytidin)  were  not  changed  in  the 
neutralized  gastric,  pancreatic  or  intestinal  juices.  The  mononu- 
cleotides (guanylic  acid  and  the  pyrimidin  nucleotides)  .were  not  af- 
fected by  gastric  or  pancreatic  juice;  but  with  intestinal  juice 
changes  were  produced  which  are  interpreted  as  showing  that  phos- 
phoric acid  was  split  off  from  the  guanosin  or  pyrimidin  complex, 
with  no  further  change  of  these  complexes.  The  action  was  much 
slower  in  the  case  of  the  pyrimidin  nucleotides  than  in  that  of  the 
guanylic  acid.  The  complex  nucleic  acids  (yeast  nucleic  acid  and 
thymus  nucleic  acid)  were  little  changed  by  gastric  or  pancreatic 
juice,  but  considerably  (rapidly  in  the  case  of  the  yeast  nucleic 
acid)  by  intestinal  juice.  The  changes  are  interpreted  as  indicat- 
ing a  cleavage  into  phosphoric  acid  and  an  organic  complex,  but  no 
reducing  substance  (carbohydrate)  was  set  free.  These  experi- 
ments seem  to  indicate  that  nucleic  acids  are  not  digested  or  ab- 
sorbed by  the  stomach,  but  that  in  the  intestine  phosphoric  acid  is 
split  off  under  the  action  of  intestinal  juice.  The  going  into  solu- 
tion which  others  had  observed  as  apparently  brought  about  by 
trypsin  may  have  been  without  this  splitting,  being  simply  such  a 
change  as  Araki  noted. 

According  to  Schaumann,  "Schittenhelm  (Arch,  f .  klin.  Med.81, 
1904,  423)  found  that  in  disease  of  the  pancreas  the  digestion  of 
nuclein  suffers,  and  he  thinks  that  the  chemical  experiments  cor- 
respond with  the  microscopical  observations.  In  considering  the 
latter,  he  points  to  the  observations  of  Schmidt,  who  showed  long 
ago  that  in  failure  of  pancreatic  ferment  unchanged  cell  nuclei  of 
the  food  are  passed  with  the  feces." 

Abderhalden  and  Schittenhelm's  (1906a)  artificial  digestions  of 
the  sodium  salt  of  a-thymonucleic  acid  with  the  juices  of  the  diges- 
tive tract,  and  with  water  extracts  of  pancreas  and  intestine, 
showed  no  determinable  change  by  the  gastric  juice,  and  with  the 


234  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

pancreatic  juice  none  which  went  far  enough  to  yield  purin  bases, 
though  there  was  some  unknown  kind  of  alteration  which  made  the 
nucleic  acid  more  readily  soluble,  and  increased  the  amount  of  di- 
alyzable  matter.  The  extracts  of  pancreas  and  intestine,  however, 
produced  an  abundance  of  purin  bases  and  a  clear  solution.  Inter- 
preted as  indications  of  the  place  in  which  cleavage  occurs  within 
the  body,  these  experiments  point  to  the  understanding  that  nucleic 
acids  do  not  undergo  deep  cleavage  by  digestive  enzymes  within  the 
intestine  but  only  in  and  beyond  the  intestinal  wall. 

EFFECTS  OF  BACTERIA  IN  THE  ALIMENTARY  TRACT 

Bacteria  in  the  digestive  tract,  especially  in  the  large  intestine, 
and  perhaps  also  such  as  are  introduced  with  the  food,  may  have 
considerable  to  do  with  the  disposition  of  nucleins.  These  micro- 
organisms show  the  power,  not  only  to  cleave  nucleins  in  the  ways 
shown  above,  but  also  to  change  amino-purins  to  oxypurins,  even  as 
far  as  uric  acid.  Doubtless  they  produce  such  effects,  as  they  re- 
main for  considerable  time  in  contact  with  the  food  material. 

It  should  be  remembered  that  these  bacteria  further  complicate 
the  studies  of  nuclein  digestion  because  the  purins  of  the  feces  may 
come  from  their  bodies,  Schittenhelm  and  Tollens  say  to  the  extent 
of  31.3  percent.  Such  purins  would  be  the  result  of  constructive 
as  well  as  destructive  activities  of  the  bacteria. 

Schittenhelm  (1903)  has  shown  that  nearly  all  of  the  nuclein 
substance  of  feces  disappears  as  they  undergo  autoputref action ;  and 
Schittenhelm  and  Schroeter  (1903a,  1903b,  1904)  showed  that  bac- 
teria may  bring  about  a  deep  cleavage  of  yeast  nucleic  acid. 

Schaumann  tells  us  that  Emmerich  and  Low  (Zeit.  f .  Hygiene 
36,  1901,  9)  found  in  pathogenic  bacteria  enzymes  which  were  ef- 
fective to  dissolve,  not  only  their  own  nucleoproteins,  but  also  those 
of  other  species  of  bacteria. 

Plenge  (1903)  showed  that  some  bacteria  have  the  power  to 
liquify  the  sodium  salt  of  a-nucleic  acid  from  thymus  and  that  the 
action  is  brought  about  by  means  of  an  enzyme. 

Vegetable  moulds  were  shown  by  Iwanoff  (1903)  to  contain  or 
produce  an  enzyme  which  completely  cleaves  nucleic  acids. 

Hahn  and  Geret  (1900)  found  evidence  of  an  enzyme  in  the  ex- 
pressed juice  of  yeast  which  has  a  cleaving  action  on  proteins,  in- 
cluding nucleoproteins,  which  is  much  like  that  of  trypsin  but  calls 
for  slightly  acid  solution.  It  frees  the  bases  and  amino-acids  di- 
rectly, without  forming  peptones,  and  but  very  little  intermediary 
albumose.  After  one  hour's  digestion  at  37°C.  the  greater  part  of 
the  phosphorus  was  converted  to  the  form  of  phosphoric  acid.  The 
enzyme  is  designated  as  yeast  endotrypsin,  and  it  is  thought  that 
it  may  play  an  important  part  in  disassimilation. 


PHOSPHORUS  METABOLISM  235 

FERMENTS  OF  INDIVIDUAL  ORGANS 

The  enzymatic  actions  studied  in  various  organs  have  included 
those  of  nucleases,  guanase,  adenase,  xanthoxidase  and  uricase.  All 
of  these  except  the  nucleases  concern  only  the  purin  fraction  of  the 
compounds.  Since  the  oxypurins,  xanthin  and  hypoxanthin,  are 
thought  to  be  absorbable,  and  the  amino-purins  not,  and  since  any 
processes  concerned  with  the  disposal  of  the  nuclein  substance  may 
be  of  importance  in  the  consideration  of  nucleins  as  foods,  we  are  in- 
cluding a  number  of  the  many  references  to  studies  of  the  location 
of  purin  enzymes,  but  we  discuss  only  those  on  nucleases,  as  having 
a  direct  bearing  on  the  freeing  of  phosphorus  from  the  molecule. 

The  most  productive  workers  in  this  field  have  been  Jones  and 
Schittenhelm  and  their  co-laborers.  References  are  listed  below. 
The  article  by  Wells  (1910)  summarizes  in  detail  the  enzymes  which 
had,  at  the  time  of  his  writing,  been  located  in  the  organs  of  dif- 
ferent animals.  Nuclease  was  found  in  all  organs  tested,  but  other 
ferments  were  rather  variously  distributed,  and  it  is  noticed  that 
species  differ  in  this  respect. 

Pighini's  (1910,1911)  optical  methods  used  in  determining  nu- 
cleic acid  cleavage  by  blood  serum  are  called  in  question  by  Amberg 
and  Jones  (1911a).  They  find  no  cleavage  of  thymus  nucleic  acid 
by  blood  sera ;  yeast  nucleic  acid,  they  say,  is  altered  in  some  way, 
and  perhaps  decomposed,  but  without  liberation  of  either  phospho- 
ric acid  or  purin  bases. 

A.  J.  Juschtschenko  (1911)  (see  also  Yushchenko,  1912)  has 
studied  the  relative  nuclease  content  of  the  blood,  muscles  and  var- 
ious organs  of  man,  dog,  horse,  cow,  rabbit,  hen  and  pike-perch, 
by  making  inorganic  phosphorus  determinations  on  uniformly  pre- 
pared solutions  containing  salt-water  extracts  of  the  organs  and 
sodium  nucleate  solution,  after  the  mixed  solution  had  stood  for  40- 
42  hours  in  a  thermostat  at  37-38°.  The  data  reported  show  the 
milligrams  of  P205  set  free  from  the  nucleate  by  one  gram  of  organ. 
The  general  conclusions  from  the  work  are  thus  expressed : 

"Liver,  kidney,  spleen,  pancreas,  and  thyroid  gland  contain  sig- 
nificant quantities  of  nuclease;  brain,  suprarenals,  lungs,  and  lym- 
phatic glands  contain  smaller  quantities;  heart,  blood,  muscle  and 
serum  are  poor  in  nuclease.  The  blood  of  dogs,  rabbits  and  cattle 
is  richer  in  nuclease  than  that  of  man.  The  livers  of  man,  horse, 
cattle,  rabbit  and  pike-perch  are  richer  in  nuclease  than  that  of  the 
dog.  In  most  organs  of  young  dogs  the  nuclease  content  is  less 
than  in  the  same  organs  of  grown  dogs.  The  organs  of  man  are  in 
general  rich  in  nuclease." 

Later  (A.  Juschtschenko,  1911),  thyroidectomy  was  found  to 
reduce  the  nuclease  content  of  the  organs  and  blood  of  young  dogs 
and  the  blood  of  rabbits. 


236  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

E.  K.  Marshall,  Jr.  (1913)  has  determined  that  the  ferments 
of  the  thymus  can  not  digest  all  of  the  nucleic  acid  of  the  gland,  but 
leave  a  portion  completely  unaltered. 

According  to  Levene  and  Medigreceanu  (1911d),  "there  exist 
in  the  organism  several  enzymes,  which  act  harmoniously,  leading 
to  the  disintegration  of  nucleic  acids.  The  function  which  hither- 
to has  been  ascribed  to  the  'nucleases'  is  in  reality  performed  by  at 
least  three  enzymes,  or  rather  three  groups  of  enzymes,  which  have 
to  be  designated  by  special  names.  The  part  of  each  enzyme  is  to 
render  the  nucleic  acid  molecule  susceptible  to  the  action  of  one  of 
the  other  enzymes,  thus  producing  successive  disintegration." 

Nucleinases  are  those  enzymes  which  cause  the  dissolution  of 
the  nucleic  acid  molecule  into  nucleotides.  Nucleinase  is  present  in 
practically  all  organs  and  in  the  pancreatic  juice.  It  is  not  present 
in  gastric  juice. 

Nucleotidases  split  nucleotides  into  phosphoric  acid  and  a  car- 
bohydrate-base complex.  They  are  present  in  the  plasma  of  all  or- 
gans, and  in  the  intestinal  juice;  they  are  not  present  in  gastric  or 
in  pancreatic  juice. 

"Also  in  regard  to  nucleotidases  the  possibility  is  not  excluded  that 
there  exists  more  than  one  enzyme  of  the  same  group,  and  this  for 

the  following  reasons [There  is]  gradation  in  the  stability 

of  the  nucleotides  towards  chemical  agencies [and] 

towards  enzymes.  Thus,  guanylic  acid  is  hydrolyzed  readily  into 
phosphoric  acid  and  guanosin  by  the  plasma  of  the  pancreatic  gland, 
whereas  there  could  not  be  ascertained  the  occurrence  of  the  anal- 
ogous cleavage  of  inosinic  acid,  of  pyrimidin  nucleotides,  nor  of  the 
complex  nucleic  acids,  through  the  action  of  the  same  plasma.  On 
the  other  hand  the  cleavage  takes  place  in  all  nucleotides  under  the 
action  of  the  extract  of  the  intestinal  mucosa." 

Nucleosidases  split  nucleosides  (purin  ribosides)  into  their  com- 
ponents. They  are  present  in  the  plasma  of  most  organs  examined 
but  not  present  in  the  plasma  of  the  pancreas,  nor  in  gastric,  pan- 
creatic or  intestinal  juice. 

"Pyrimidin  Complexes.  Comparatively  little  information  is 
obtained  regarding  the  mechanism  through  which  these  substances 
undergo  disintegration  in  the  animal  organism.  The  only  evidence 
of  the  possible  existence  of  enzymes  bringing  about  cleavage  of  these 
complexes  may  be  found  in  the  older  observation  of  Levene,  that  in 
course  of  prolonged  autolysis  of  organs,  free  pyrimidins  are 
formed." 


PHOSPHORUS  METABOLISM  237 

A  viscosity  method  has  been  used  by  de  la  Blanchardiere  (1913) 
in  investigation  of  nucleolytic  enzyme  activity.  From  such  obser- 
vations, made  with  organ  extracts  and  pancreatic  secretion,  it  is 
concluded  that  nuclease  is  not  identical  with  any  proteolytic  en- 
zyme ;  also  that  the  processes  of  liquefying  and  of  cleaving  the  nu- 
cleic acid  are  distinct,  the  work  either  of  two  different  nucleases  or 
of  two  different  groups  of  the  enzyme  molecule. 

References  on  organ  tests:  Horbaczewski,  1889,  1891a,  1891b,  1893, 
Schwiening,  1894;  Spitzer,  1899;  Kutscher,  1901;  Araki,  1903a;  Jones,  1904a, 
1904b,  1904c;  Jones  and  Partridge,  1904;  Schittenhelm,  1904a,  1904b,  1905a, 
1905b;  Jones  and  Winternitz,  1905;  Jones,  1905;  Schenck,  1905;  Burian,  1905; 
Sachs,  1905;  Bloch,  Bruno,  1906;  Abderhalden  and  Schittenhelm — resume  of 
literature— 1906a;  Schittenhelm  and  Schmidt,  1906,  1907a,  1907b;  Jones  and 
Austrian,  1906,  1907a,  1907b;  Arinkin,  1907;  Mendel,  L.  B.,  and  Mitchell,  1907-8; 
Jones,  1908;  Steudel,  1908a,  1908b;  Kiinzel  and  Schittenhelm,  1908,  1909;  Schit- 
tenhelm, 1908,  1909a,  1909b;  Wells  and  Corper,  1909;  Leonard  and  Jones,  1909; 
Rohde  and  Jones,  1909;  Miller  and  Jones,  1909;  Straughn  and  Jones,  1909; 
Winternitz  and  Jones,  1909;  Jones,  1910;  Wells,  1910;  Pighini,  1910,  1911; 
Amberg  and  Jones,  1911a,  1911b;  Jones,  1911a,  1911b;  A.  J.  Juschtschenko, 
1911;  A.  Juschtschenko,  1911;  Medigreceanu,  1911;  Levene  and  Medigreceanu, ' 
1911b,  1911d;  Mihari,  1911;  Corper,  1912;  Schittenhelm  and  Wiener,  1912; 
Yushchenko,  1912;  Levene  and  LaForge,  1913;  de  la  Blanchardiere,  1913;  Mar- 
shall, 1913. 

ANIMAL  EXPERIMENTS  ON  THE   DIGESTION   OF   NUCLEOPROTEINS 

AND  NUCLEIC  ACIDS 

Loewi  (1900,  1900-01)  conducted  metabolism  experiments, 
mostly  with  human  subjects,  in  which  observations  were  made  of 
the  effects  on  excretion  of  adding  various  nuclein  substances  to  a 
mixed  diet.  In  the  work  reported  in  the  first  paper  thymus  gland 
was  substituted  for  beef  in  an  otherwise  constant  mixed  diet  with  a 
subject  suffering  from  myelogenic  leukaemia,  and  with  a  normal 
subject;  the  effect  of  adding  pancreas  to  a  constant  mixed  diet  was 
also  studied;  and  a  search  was  made  for  allantoin  in  human  urine 
after  the  ingestion  of  pancreas,  desiccated  pancreas  or  thymus.  It 
was  concluded  (in  part)  that :  The  excretion  of  uric  acid  in  leukae- 
mics  is  not  different  from  that  in  normal  individuals;  there  is  di- 
uresis. The  ratio  of  uric  acid  to  P205  does  not  increase  with  math- 
ematical precision  following  thymus  feeding.  Both  the  items  are 
increased  by  the  gland.  Increased  katabolism  does  not  occur, 
since  the  nitrogen  figures  return  to  normal  shortly  after  discontin- 
uing thymus  feeding.  No  allantoin  could  be  found  in  the  samples 
of  human  urine  examined. 

The  work  reported  in  the  second  paper  includes  similar  tests 
with  salmon  nuclein,  pancreas  nuclein,  yeast  nuclein,  nucleic  acid 
and  "Nutrose"  (a  neutral  sodium  casein  preparation  from  milk) ; 
also  thymus  was  fed  to  a  dog  having  ligated  pancreatic  ducts,  and 
P205  was  estimated  in  the  excreta ;  and  the  uric  acid  excretion  in  the 
urine  was  compared  in  three  subjects  on  the  same  ration. 


238 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


02 

£ 

O 

■— !03CO^OC3}CMlOi— 1 

HH 

O-HOOOOO-HO 

fa 

o 

Ph 

+++++++++ 

fa 

3 
Ed 

M 

53 

CNCNO'«^'-H**t<CDaGa3 

omo-Hp«OHO 

i +++T++++ 

>* 

fa 

O 

G 

-^^HCMOilOCMCOlOlO 

fa 

w 

Pm 

cotoco^co-^coioco 

H 

r: 

O 

H 

t—     NMto     co 

fa 

(XI 

mooioco  —  io-jicdcm 

55 

t^lO-*(MI>r~^<00O 

mi^mr--mcomcoTO 

fa 

<J 

W. 

O 
Pm 

< 

05-^r~00C5O05t^CN 

OrtO-IHHOHH 

o 

0) 

£ 

u 

t— 1 

&H 

fa 

fa 

COCM^-COC<lCCCD-*t<m 

53 

O5COiniaOjCO0Cr-O5 

T-IrHt-HC^oj-Hi-5i-<--H 

£ 

3 
P. 
■p 

3 

•a 

o 

cococoomcocoooTfi 

> 

t-i 

P 

inoeoiot^ocoxt- 

OHOHOHOOO 

«• 

oogocgocMint^cio 
oooocoiNinoot^'j 

ANCES,WITH 

900-01)— Gram 

PERIMENT  I 

P* 

s 

o 

cocoo5-siomE^oo"»i 

■^ir*-^oiocM'**<r^co 

a. 

CMCOC^COOJCQCNCOOJ 

NONNCDOCOOO 

far}  K 

53 

C^CCOOjCO-^OCMt— IO 

«3       « 

coioco-*fcomcoioco 

SI 

|Q 

o 

inoioccoiNttJHt-. 

w 

Ah 

coi— coirjcomcor^co 

o 

w 

fa 

QQ 

coiococxcor^iococN 

IOOOIOCOIOt— it^t^OS 

& 

O 

toomoomasiraoom 

fa 

fl 

p 

•S    '1    .9    * 

Jz; 

■3     §    JB     o 

-< 

3    c    y    rt 

C                   3 

-     2     3     H 

fa 

U.5        H        £        3 

2  ts-d  e-a  g,T3  c-d 

•c 

a 

o 

o 

•d    .rtCrt^rtcrt 

BO  4->    M+jg+jCM  jj 

fa 

H 

»— i 

■Ml        CM       -4-        1 

K+    +    Z    -o 

A    e    *    -d 

•d       Ed      ■§       c 
3      -d       5       rt 

a     5    *    m 

* 

fa 

HH 

<l3 

Q 
fa 

O 

c>~ 

>> 

■"Ji^HCOi-KMi-ICOi— ICN 

53° 

•e 

fa 

o 

MK£>t>£K£K 

Cm 

!>> 

CO 
t—i— IO 

rHCNieO 
++  + 

CD 

innoo 
ocmcm 

+++ 

m-^ti-n 
r-r^co" 

COCOIO 
lO-WCO 
ENIOCO 

03C33CN 
TJ1T}<CO 
CO  COO) 

mco-H< 

00  coo 

■-ICMO 

H 

H 

M 

EH 

53 
W 

9 

H 

coinos 
ast^-co 

CO  CO  CO 

M 
W 

W 

OiCOH 

cooco 

OCOCD 

00  CO  CO 

i— ICO»-l 

or-o 

t^CMt~ 

cno'oi 

1- ICN^H 

3} 
K] 

jb 
"o 

3  ' 

H 

"SI'S 
•3  °3 

CeO 

+ 

■d 

Ed 

•d 

3 
rt 
+j 

•«<COCM 

HHH 
t-H 

+++ 


r~r-oo 


eooco 
corded 


00O3  00 

o'o'o 


00t~O5 

cmcn'o) 


•— 10503 


88S1 

COTfi"* 


IOIOCO 
t^COOO 


<p  n-d 

t-i  aj  i-t 

Ed  b,  rt 

•d  .-a 

3  a  3 
rt  3  rt 

CO 

+ 


I      I 


0CTt<O5 


00  CM 
IOOCO 
i— I  (Mi— ( 

E-^a>od 


oaco-*^ 

CO  CO  CO 


IOC5CO 

t^oooo 


■d  E-d 

i-    :i  i- 

•d^-d 

Ed  i    Ed 


i*     SS 


tic  R. 


*"*    s 


fl> 


X 

o 

P. 

R 

p 

H 

bo 

fcj 

3 

HO" 


09 

■3 

■d-a 

a 

fH-a 

H 

cS 

3 

<3 

3 

C3 

o 

- 

rH 

|M 

uO 

S 

o 

•l)t-K    . 
-       04  W*S 

"o     N  <a 

om*<s  S 

-04 


01 


-3   3  b-""    « 
§  o  S  g  g 

E0  O^    O    (D 

g'co    -  p.  a, 

n-d  °  ° 


osa. 


8  «OEJ 

o  *J  3  -zTS 
COOQM 


ca 


PHOSPHORUS  METABOLISM  239 

Conclusions  (in  part).  "The  nucleins  of  the  food  are  in  part 
cleaved  in  the  intestine ;  the  P205  of  the  cleaved  fraction  passes  into 
the  feces,  the  nitrogenous  fraction  is  resorbed.  The  larger  portion 
uncleaved  is  resorbed  in  to  to,  whereby  the  P2Og  remains  in  organic 
combination." 

"It  is  possible  by  nuclein  feeding  to  cause  a  retention  of  nitro- 
gen and  phosphorus  in  the  body  in  the  same  proportions  that  these 
substances  are  present  in  the  ingested  nuclein." 

"Nuclein  superposition,  under  certain  conditions,  encourages  ni- 
trogen as  well  as  phosphorus  retention." 

"Excepting  the  uric  acid,  no  other  specific  nitrogen-  or  phospho- 
rus-containing end-products  of  nuclein  metabolism  occur  in  appreci- 
able quantities  in  human  urine." 

"Uric  acid  excretion  normally  is  dependent  only  upon  diet." 

Mochizuki  (1901)  judged  from  an  increase  of  nitrogen,  uric 
acid  and  phosphorus  in  the  urine  that  the  proteins  of  a  thymus 
broth  were  readily  absorbed  through  the  colon  when  introduced 
per  rectum. 

Sweet  and  Levene  (1907)  studied  nuclein  metabolism  in  a  dog 
with  an  Eck  fistula,  that  is,  an  anastomosis  between  the  portal  vein 
and  the  inferior  vena  cava,  an  operation  which  cuts  off  the  activity 
of  the  liver  in  oxidizing  uric  acid  to  urea.  The  experiments  were 
made  in  three-day  periods  after  several  weeks'  feeding  on  the  purin- 
free  diet  which  was  used  as  the  basal  ration  in  the  experimental 
periods.  Observations  were  made  (1)  with  the  purin-free  diet,  (2) 
on  a  diet  containing  the  same  amount  of  nitrogen  but  partly  in  the 
form  of  nucleoprotein,  adenin  sulphate,  sodium  nucleate  or  thymin ; 
also  (3)  on  a  diet  of  adequate  calorific  value,  but  low  in  nitrogen, 
and  finally,  (4)  when  no  food  was  taken.  The  purin-free  diet  was 
composed  of  plasmon,  cracker  dust  and  lard;  and  the  low-protein 
diet  was  composed  of  cane  sugar,  cracker  dust  and  lard. 

Nitrogen  equilibrium  was  maintained  on  the  non-purin  diet,  the 
uric  acid  output  being  higher  than  normal.  Nucleoprotein  feeding 
slightly  increased  the  uric  acid  output.  When  1.0  gm.  adenin  was 
fed  there  was  an  increase  in  uric  acid  corresponding  to  44  percent  of 
the  base  given.  Ten  grams  of  sodium  nucleate  increased  the  uric 
acid  by  more  than  40  percent  of  the  purin  given,  the  phosphorus 
outgo  also  showing  increase.  Thymin  was  eliminated  mostly  as 
such.  Thymin  was  not  observed  in  the  urine  on  either  nuclein-free 
or  nucleic  acid  diets.  Nucleic  acid,  therefore,  is  not  completely  dis- 
integrated, or  its  decomposition  products  are  slowly  formed,  in  small 
quantities,  and  immediately  destroyed.  An  increased  uric  acid  out- 
go on  the  low-protein  diet  indicated  increased  cell  katabolism,  and  a 


240  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

subsequent  decreased  output  showed  an  adjustment  of  the  metab- 
olism to  the  continued  low-protein  diet,  this  elimination  being  but 
slightly  increased  in  the  starvation  period  following. 

Abderhalden,  London  and  Schittenhelm  (1909)  also  made  use 
of  an  Eck  fistula  in  a  dog  in  studying  the  influence  of  the  liver  in 
nuclein  metabolism.  From  urine  observations  in  connection  with 
nucleic  acid  feeding  it  was  concluded  that  the  digestion,  desamidiza- 
tion  and  oxidation  of  the  nucleic  acid  proceeded  nearly  as  usual ;  the 
uric  acid  was  not  transformed  into  allantoin  to  quite  the  extent  that 
Schittenhelm  had  found  usual  with  normal  dogs.  Both  uric  acid 
and  phosphorus  in  the  urine  rose  when  the  sodium  nucleate  (and 
starch)  was  added  to  a  bread  and  milk  diet. 

In  another  experiment  with  dogs  (Schittenhelm,  1909a)  sodium 
nucleate  was  either  fed,  or  introduced  with  a  probang,  (and  in  one 
case  intravenously)  and  the  urine  and  feces  were  examined.  Obser- 
vations of  this  kind  were  made  with  normal  dogs,  starved  dogs,  and 
dogs  under  the  influence  of  alcohol.  The  nitrogen  elimination  indi- 
cated that  the  nucleic  acid  when  introduced  by  the  mouth,  was  com- 
pletely absorbed,  and  then  its  purins  were  almost  wholly  eliminated 
as  allantoin,  though  all  the  forms  of  purin  nitrogen  in  the  urine  were 
appreciably  increased.  Injected  salt  was  not  eliminated  in  the  same 
way  as  that  fed ;  the  nitrogen  eliminated  was  increased  by  an 
amount  much  more  than  that  added,  and  considerably  more  of  it 
was  in  the  form  of  purin  bases.  Vomiting  sometimes  followed  the 
introduction  by  probang.  The  influence  of  the  chronic  alcoholism 
was  evident  in  a  delayed  elimination,  probably  delayed  metabolism 
of  the  nucleic  acid,  purins  showing  in  the  allantoin  output,  just  as  in 
human  alcohol  experiments  uric  acid  elimination  is  delayed.  Purin 
excretion  observations  were  made  by  Schittenhelm  and  Bendix 
(1904,  1905)  after  feeding  or  injecting  nucleic  acid  in  rabbits. 

F.  Frank  and  Schittenhelm  (1909)  also  report  nitrogen  and 
phosphorus  retention  experiments  with  three  human  subjects  in 
which,  to  a  constant  diet  of  cereals,  milk,  eggs,  butter  and  fruit 
(purin-free)  there  was  added,  on  the  experiment  days,  10  grams  of 
the  sodium  salt  of  thymus  nucleic  acid.  Determinations  were  made 
of  total  nitrogen,  urea,-  uric  acid-,  and  purin  base-nitrogen,  as 
shown  in  the  table.  Patient  III  had  a  stricture  of  the  oesophagus 
and  was  fed  by  stomach  fistula.  The  authors  concluded  that  there 
was  a  prompt  working  over  of  the  nucleic  acid,  with  quantitative 
elimination  of  the  end-products  after  only  one  day.  The  purin  bases 
of  the  nucleic  acid  appeared  mainly  in  the  form  of  urea,  but  little 
as  uric  acid,  and  but  a  very  small  part  as  purin  bases. 


PHOSPHORUS  METABOLISM 


241 


fa 
O 

bq 

OS  fc 
o  < 
fa  S 

fa  & 


o 

o 

I— I 
H 


fa 

fa 
O 
O 
fa 
H 


fa 

fa  A 

fa      OS 

a  5 

fa  M 

2  -a 

H 

<u 


S 
oS 

s 

c3 
u 
fa 


IB 

o 

o 

KlOt^ 

■^flCDCO 

0305 
QOtDO 
CO  CO  CO 

TH010 
COCOH 

d.  a 

0*0  0 

00 -*H 

loom 

t— (C\lrH 
OOO 

CO  CM  CO 

omo 
ooo 

OOO 

COOS 

oaco'c<i 

000050 

cooor- 

COrHrH 

SOrH 
COOJ-* 

OOO 

4) 

« 
'C 

o 

COCOtM 
TJ100CO 

oaoJoa 

eococi 

N00 

CO  OHO 
COCM'fl 

i-HCOi-H 

-*T*<,-H 
OOO 
OOO 

H1O00 

HHO 
OOO 

OOO 

lOTtlt~ 
OHO 
OOO 

OOO 

OlOH 

o-*c<i 
o'oo 

,    C0CO00 
0210  05 
OHO 

OOO 

cot^t^ 

COC0  03 
OINO 

oo"o 

OOSCD 

in-*  oo 

00  05  00 

CO00IM 
t^-OOCO 

0500 

CO 

ocom 

tOh-H 

iraio'io 

"iS 
■gfc 

THC01O 
OOH 

OHO 

oeo<M 
oqcoTti 

rHOaH 

mt»ira 

iomo 

CO  CD  CO 

■a 

o 
o 

Eh 

'o 

_o 
'53 

o 
3 

o 
£  - 

d  u 

•  OO     • 

•  o     • 

•  CO      • 

'  -o    • 

.00    . 
•  o     • 

.50      . 
•O      - 

.00     •     . 
•  o     • 

■  CO      • 

:o   : 

in 

OIO<D 
CO-^HCO 

lOOlO 
co-^ico 

lOCo'lO 

t»<N-H1 

i-HCOCO 

odojoi 

Body 
weight 

of 
subject 

Kg. 

HOH 
!CS3 

coooco 

IOIOCO 

00  CO  CD 

ioio'io 

CM  CO  CM 

C 

c 

c 

Length 

of 
period 

Days 

-*>nco 

^ 

CO  CO  CO 

+> 

e 

0) 

.S 

u 

V 
P. 

■d 

.8 
'C 
<u 

p. 
d 
S 

"d-d  "S 
.g.g-g 

2  »$ 

O  K*£ 

o  o.S 
'u'u  u 

O   X"" 

M 
M 

d'-d'S 
o  o.S 

'§'§& 
ftp,* 

ox"-; 

M 

M 

< 

242 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


This  finding  of  urea  as  the  chief  end-product  of  the  purin  nitro- 
gen of  nucleic  acid  in  man  is  not  in  harmony  with  the  view  held  by 
many  that  uric  acid  must  be  the  end-product  because  in  man  no  uri- 
case  is  found.  Dohrn  (1913)  reports  one  experiment  in  which' the 
urea  was  not  uniformly  increased  by  nucleic  acid  intake.  In  his  ob- 
servations both  nitrogen  and  phosphoric  acid  elimination  were  in- 
creased beyond  the  intake. 

Marfori  (1908b)  notes  that,  by  administration  of  nucleins,  not 
only  all  the  phosphorus  of  the  nuclein  appears  in  the  urine,  but  an 
excretion  greater  than  corresponds  to  the  same.  The  mass  of  the 
phosphorus  found  in  the  feces  is  increased.  Nuclein  or  nucleic  acid 
given  by  mouth  to  dogs  is  absorbed,  and  excreted  in  the  urine  in  the 
form  of  phosphates.  Nucleoproteins  are  consequently  not  in  con- 
dition to  offer  assimilable  phosphorus  to  the  organism.  This  ob- 
servation is  not  in  accord  with  prevailing  opinion. 

Oeri  (1909),  from  a  brief  experiment  on  himself,  aged  27,  con- 
cluded that  the  phosphorus  of  nucleate  is  set  free  in  the  form  of 
phosphoric  acid  in  the  intestine,  and  that  the  path  of  its  elimination 
is  then  determined  largely  by  the  presence  or  absence  of  calcium.  If 
calcium  is  present,  calcium  phosphate  forms  and  is  excreted  by  the 
intestine ;  but  if  calcium  is  not  available,  the  phosphoric  acid  is  ex- 
creted by  the  kidneys.  The  nucleate  was  taken  in  a  single  dose  of 
12.99  gm.  of  the  sodium  salt;  conditions,  therefore,  were  not  favor- 
able to  observations  on  nitrogen  and  phosphorus  retention  from  nu- 
clein. 

Sokolov  (1912)  is  said  to  show  that  sodium  nucleate  is  not  as- 
similated by  the  aged,  but  that  its  assimilation  is  marked  in  young 
individuals. 

From  a  study  of  nuclein  metabolism  in  a  pig,  a  part  of  the  re- 
sults of  Schittenhelm  (1910)  are  as  follows: 


AVERAGE  DAILY  NITROGEN  AND  PHOSPHORUS  BALANCES  WITH  A 
PIG,  AS  AFFECTED  BY  NUCLEIC  ACID— Grams 


5  °  £ 

Food 
N 

Food 
P2O5 

Urine 

Feces 

N 

Uric 
acid 

N 

Purin 
N 

Allan- 
toin 

N 

P2O5 

N 

Purin 

N 

P2O5 

Ration. 

5 
3 

3 
5 

5 

5 

8.7 
9.49 

8.7 
14.07 

16.12 

14.07 

3.80 
4.40 

3.80 
5.30 

7.65 

5.30 

5.84 
6.57 

6.30 
5.20 

6.38 

6.40 

0.005 
0.009 

0.009 
0.006 

0.010 

0.009 

0.016 
0.029. 

0.018 
0.027 

0.054 

0.025 

0.236 
0.575 

0.255 
0.278 

0.973 

0.374 

1.20 
1.63 

1.23 

1.07 

3.32 

0.265 
0.682 

0.193 
0.550 

0.210 

0.295 

0.002 
0.011 

0.009 
0.037 

0.011 

0.016 

0.39 
0.695 

0.191 
0.45 

0.20 

0.31 

1500  cc  milk. 
Same  plus  20  g-m. 

nucleic  acid  from 

yeast. 
1500  cc.  milk. 
1500  cc.    milk;   300 

g-m.  wheat  flour. 

Same  plus  14  arm. 

yeast  nucleic  acid. 

1500  gm.  milk;  300 

gm.  wheat  flour. 

PHOSPHORUS  METABOLISM  243 

In  the  first  experiment  the  pig  was  8  weeks  old ;  in  the  second, 
4  months  old. '  In  the  first  there  was  no  gain  in  weight ;  in  the  sec- 
ond the  pig  increased  in  weight  from  25.4  to  31.6  kg.,  or  413  gm.  per 
day.  The  author  concludes  that  nucleic  acid  fed  to  swine  is  easily- 
absorbed  and  completely  broken  down ;  the  purin  portion  appears  as 
the  end-product  allantoin,  while  the  intermediate  products,  uric  acid 
and  purin  bases,  are  eliminated  as  such  only  in  very  small  part, 
about  1-5  percent.  In  the  second  experiment,  and  in  another  in 
which  there  was  gain  in  weight,  the  elimination  of  nucleic  acid  was 
not  quantitative,  which  indicates  that  the  nucleic  acid  of  the  food 
can  be  retained  and  utilized.  In  other  experiments  the  author 
showed  that  allantoin,  either  fed  or  injected,  was  excreted  un- 
changed. 

Meier's  (1910)  work  also  shows  that  in  hogs  nucleic  acid  is 
largely  absorbed,  and  the  purin  fraction  appears  as  allantoin  in  the 
urine,  with  only  small  amounts  of  uric  acid  and  purin  bases. 

Levene  and  Medigreceanu  (1911a),  after  pointing  out  some  of 
the  difficulties  and  uncertainties  in  most  of  the  previous  attempts 
to  learn  of  the  exact  processes  and  paths  by  which  nucleins  are 
transformed  to  the  end-products  uric  acid,  allantoin  and  urea,  report 
data  of  their  own,  obtained  from  a  dog,  to  which  were  fed  various 
products  from  the  partial  breaking  down  of  the  acid,  as  well  as  nu- 
cleic acid  itself.  An  attempt  was  made  to  maintain  the  animal 
in  nitrogenous  equilibrium  between  experiments.  No  new  experi- 
ments were  performed  before  the  animal  returned  to  its  normal  con- 
dition. The  substances  employed  were  allantoin,  hypoxanthin,  ino- 
sin,  nucleic  acid,  and  thymus  gland.  The  urine  was  analyzed  for  the 
following  substances;  total  nitrogen,  uric  acid,  purin  bases,  am- 
monia, amino  nitrogen  and  allantoin.  Percentage  transformation 
of  the  fed  purin  was  calculated  on  the  basis  of  nitrogen  eliminated 
in  the  feeding  experiments  in  excess  of  the  nitrogen  output  in  the 
normal  periods. 

The  authors  conclude  that  the  urinary  constituents  derived  dur- 
ing 24  hours  from  the  purin  nitrogen  of  the  nucleic  acid  adminis- 
tered were  85  percent  allantoin  and  15  percent  urea;  and  that  the 
urinary  constituents  excreted  in  24  hours  from  the  purin  nitrogen 
of  thymus  were  17  percent  allantoin,  5  percent  uric  acid  and  78  per- 
cent urea.  There  was  no  increase  in  the  amino  nitrogen  output  af- 
ter any  one  of  these  experiments. 

"From  the  results  of  these  experiments  it  is  apparent  that  the 
highest  proportion  of  allantoin  output  follows  the  administration  of 
nucleic  acid  and  of  hypoxanthin;  the  proportion  is  lower  after  the 


244  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

administration  of  inosin.  Thus  it  seems  possible  that  the  first  step 
in  the  disintegration  of  nucleic  acid  in  the  organism  is  the  libera- 
tion of  purins  and  not  of  inosin." 

London  (1909)  has  presented  unique  evidence  as  to  the  nature 
and  extent  of  digestion  in  the  several  portions  of  the  alimentary- 
tract.  These  several  regions  were  studied  independently,  by  means 
of  fistulae,  at  various  points  in  the  intestine.  Experiments  were 
conducted  on  four  dogs  with  nucleoprotein  from  horse  liver.  London 
concluded  that  about  two-thirds  of  the  nucleoprotein  goes  into  solu- 
tion in  the  stomach,  but  with  the  splitting  off  of  only  2-3  percent  of 
the  phosphorus,  the  dissolved  substance  giving  the  reactions  of  nu- 
cleic acid.  In  the  intestine  nucleic  acid  is  decomposed,  the  prin- 
cipal part  in  the  digestive  process  being  played  by  the  intestinal  se- 
cretion. 

By  further  use  of  London's  fistula  method,  London  and  Schit- 
tenhelm  (1910)  watched  the  digestion  t)f  nucleic  acids  in  dogs  hav- 
ing fistulae  in  the  duodenum,  the  jejunum,  the  ileum  or  the  ileo-coe- 
cal  region.  In  one  series  of  experiments  the  dogs  were  fed  milk, 
white  bread  and  the  sodium  salt  of  yeast  nucleic  acid ;  in  two  other 
series,  white  bread  and  the  sodium  salt  of  thymus  nucleic  acid ;  in  a 
fourth,  the  sodium  salt  of  thymus  nucleic  acid  with  water  only ;  and 
in  a  fifth — a  control  experiment — they  were  fed  bread  and  milk  only. 
From  this  work  it  appears  that  nucleic  acids  are  neither  changed 
nor  absorbed  in  the  stomach ;  but  in  the  intestine  there  is  a  change 
by  which  a  small  part  seems  to  be  split,  with  the  formation  of  free 
purin  bases,  while  a  larger  part  is  so  changed  that  dialyzable  cleav- 
age products  are  produced  which  still  contain  organically  combined 
purin  bases.  To  determine  whether  the  digestion  katabolism  corre- 
sponds with  the  hydrolytic  cleavage  which  Levene  has  observed, 
Levene's  method  of  fractioning  the  lead  acetate  solution  was  fol- 
lowed, and  in  that  way  it  was  shown  that  a  change  does  take  place 
by  which  the  amount  of  unaltered  nucleic  acid  becomes  less  the  near- 
er the  fistula  is  to  the  lower  ileum,  and  products  are  formed  which 
are  rich  in  purin  bases.  According  to  Levene's  interpretation, 
these  might  be  either  a  nucleoside  (e.  g.,  guanosin),  or  a  mononu- 
cleotide (e.  g.,  guanylic  acid),  or  a  mixture  of  the  two. 

The  absorption  of  nucleic  acid  or  its  cleavage  products  is  found 
to  take  place  mainly  in  the  lower  parts  of  the  intestine,  the  lower 
jejunum  and  the  ileum,  as  indicated  by  these  cleavages  observed. 
Since  the  amount  of  free  bases  found  among  these  digestion  prod- 
ucts is  very  small,  it  would  appear  that  complete  cleavage  is  not  nec- 
essary for  absorption.  The  control  experiment  showed  th&t  the 
body  does  not  give  off  any  purin  bases  in  the  digestive  juices,  as  has 
repeatedly  been  affirmed. 


PHOSPHORUS  METABOLISM  243 

Later  communications  from  these  authors  (London,  Schitten- 
helm  and  Wiener,  1911,1912)  show  that  undoubtedly  guanosin  is 
among  the  cleavage  products  formed  in  the  intestine,  and  probably 
also  guanylic  acid  and  aclenosin,  and  that  the  active  agent  in  the 
process  is  not  the  pancreatic  juice,  but  the  intestinal  juice.  This 
last  point  is  learned  both  from  artificial  digestions  with  the  juices, 
and  from  observations,  by  the  fistula  method,  in  which  the  normal 
dog  was  compared  with  (1)  a  dog  having  no  stomach,  (2)  a  dog  with 
the  pancreatic  juice  shut  off  from  the  intestine,  and  (3)  a  dog  with- 
out a  pancreas. 

From  this  series  of  investigations,  then,  we  are  given  the  fol- 
lowing understanding  of  the  digestion  of  nucleic  acids :  If  a  polynu- 
cleic  acid  be  ingested,  it  undergoes  little  or  no  digestion  or  absorp- 
tion in  the  stomach;  but  in  the  intestine  the  intestinal  juice  first 
breaks  it  into  mononucleotides,  without  splitting  off  any  phosphoric 
acid,  and  then  the  phosphoric  acid  splits  off  from  the  mononucleo- 
tide, forming  a  mononucleoside,  which  is  a  combination  of  purin 
base  or  pyrimidin  base  and  carbohydrate.  This  process  takes  place 
during  passage  through  the  small  intestine,  the  products  apparently 
being  absorbed  in  states  of  incomplete  cleavage.  The  various  or- 
gans and  tissues  of  the  body  apparently  have  the  power  to  cleave 
further  and  to  transform  the  purin  complexes. 

Mayesima  (1913)  found  that  a  dog  with  the  anterior  90  percent 
of  the  small  intestine  removed  was  able  efficiently  to  absorb  the  ni- 
trogen and  phosphorus  of  yeast  nucleic  acid.  Assuming  that  the 
normal  region  of  absorption  of  nucleic  acid  is  the  lower  part  of  the 
small  intestine  emphasis  might  be  put,  in  the  interpretation  of  this 
observation,  on  the  activity  of  the  remaining  10  percent  of  the  small 
intestine  or  on  a  compensatory  absorptive  activity  of  the  large  in- 
testine. 

ACCOMPANIMENTS  OF  THE  INGESTION  OR  INJECTION  OF  NUCLEAR 

MATERIAL 

High  Purin  Excretion.  For  considerations  of  the  amount  and 
origin  of  the  uric  acid  of  the  urine,  especially  as  related  to  nucleins, 
see  the  extensive  discussions  and  reviews  of  literature  the  citations 
to  which  follow: 

Burian  and  Schur  (1900,  1901,  1903);  Wiener:  Die  Hamsaure;  Ergebnisse 
der  Physiol.  I  (1902),  Abt.  I;  Mendel,  L.  B.  (1906);  Brugsch  and  Schittenhelm 
(1907a,  b,  c,  d,  e,  f,  1908-9,  1910)  and  Brugsch  (1909). 

Briefer  discussions  or  reports  of  experimental  evidence  bearing 
on  this  question,  which  have  come  into  this  research  are  the  follow- 
ing: 


246  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Salomon,  1878;  A.  Kossel,  1882;  Horbaczewski,  1889,  1891a,  1891b,  1893; 
Weintraud,  1895a,  1895b,  1900;  Kuehnau,  1895;  Hess  and  Schmoll,  1896;  Mayer, 
Paul,  1896;  Camerer,  W.,  Sr.,  1896;  Umber,  1896;  Jerome,  1897-8,  1899;  Milroy 
and  Malcolm,  1898;  Minkowski,  1898;  Loewi,  1900,  1900-01;  Kaufmann  and 
Mohr,  1902;  Mendel,  L.  B.,  Underhill  and  White,  1903;  Schittenhelm  and  Ben- 
dix,  1904,  1905;  Folin,  1905;  Macleod  and  Haskins,  1906;  Sweet  and  Levene, 
1907;  Schittenhelm,  1907;  Brugsch,  i908;  Abderhalden,  London  and  Schitten- 
helm, 1909;  Schittenhelm  and  Wiener,  1909;  Scaffidi,  1909b;  F.  Frank  and 
Schittenhelm,  1909;  Schittenhelm,  1909a,  1910;  Meier,  1910;  Fromherz,  1911; 
Smetanka,  1911;  Levene  and  Medigreceanu,  1911a. 

Horbaczewski  (1889,  1891a,  1891b,  1893)  first  showed  that  the 
action  of  blood  on  pulp  of  spleen,  and  of  several  other  organs,  re- 
sults in  a  formation  of  uric  acid,  and  he  held  that  normally  uric  acid 
formation  in  mammalia  is  due  to  the  action  of  living  blood  on  the 
lymphatic  elements,  particularly  the  leucocytes.  He  considered 
the  various  causes  which  increase  the  uric  acid  of  the  urine,  includ- 
ing nuclein  ingestion,  and  noted  that  nearly  every  one  is  accompan- 
ied also  by  high  leucocytosis ;  and  it  was  his  idea  that  the  increased 
uric  -acid  came  as  an  indirect  effect  of  these  agents,  they  bringing 
about  a  high  production  of  leucocytes  which  must  be  followed  by 
much  destruction  of  leucocytes,  and  the  elimination  of  their  purins 
in  that  form. 

A.  Kossel  (1882)  suggested  that  there  might  naturally  be  con- 
nection between  the  nucleins  and  uric  acid  output,  and  it  is  now  gen- 
erally recognized  that  nucleins  of  the  food  contribute  directly  to  the 
purins  of  the  urine ;  and  accordingly  the  urinary  purin  is  spoken  of 
as  including  two  portions ;  endogenous  purins,  arising  from  the  leu- 
cocytes and  from  the  nuclei  of  tissue  cells,  and  exogenous  purins, 
arising  from  nuclear  material  introduced  into  the  body.  The  at- 
tempts of  Burian  and  Schur  to  differentiate  these  two  portions,  to 
determine  the  amounts  due  to  each,  and  to  establish  some  laws  gov- 
erning one  or  the  other  factor,  have  had  considerable  influence  on 
the  later  thought. 

Burian  and  Schur  (1900,  1901,  1903)  compute  the  endogenous 
urinary  purins  as  the  amount  eliminated  on  a  diet  which  supplies 
the  nitrogen  requirement  but  which  is  practically  free  from  purins. 
The  amount  so  determined  is  used  also  as  a  measure  of  the  exog- 
enous purins  from  a  diet  containing  purins,  Burian  and  Schur  hold- 
ing that  the  endogenous  purin  maintains  a  constant  value  for  the 
same  individual  under  the  same  conditions  of  living,  but  that  it  dif- 
fers for  different  individuals.  On  this  basis  the  exogenous  purins 
are  considered  as  originating  entirely  from  such  bodies  contained  in 
the  food ;  and  that  portion  of  the  food  purins  which  appears  as  uric 
acid  in  the  urine  is,  for  human  subjects,  dependent  on  the  nature  of 
the  food  rather  than  upon  individual  variation.      "Factors"  are  de- 


PHOSPHORUS  METABOLISM  247 

veloped  for  different  species,  by  which  the  amount  of  uric  acid  elim- 
inated during  24  hours  may  be  multiplied  to  make  an  approximate 
determination  of  that  present  in  the  blood.  Some  of  the  factors  giv- 
en are,  for  man,  2 ;  for  rabbit,  6 ;  for  carnivora,  20-30. 

Some  of  the  observers  do  not  agree  with  Burian  and  Schur  as  to 
the  constancy  of  endogenous  .purin  for  the  same  individual,  or  as  to 
the  independence  of  the  exogenous  purin  of  personal  variation. 

It  must  be  remembered  that  in  this  relation  purin  includes  all 
the  alloxuric  bodies  as  well  as  uric  acid,  that  is,  all  those  compounds 
constructed  with  the  group  C5N4  as  a  nucleus.  Both  base  and  acid 
are  increased  in  human  urine  by  nuclein  in  the  food.  In  dogs,  cats, 
pigs  and  some,  other  animals  the  increase  appears  mostly  as  allan- 
toic 

The  endogenous  purin  may  be  increased  by  whatever  body 
processes  are  accompanied  by  a  great  tearing  down  of  nucleated 
cells,  such,  for  example,  as  excessive  strain  on  the  digestive  organs, 
and  the  destruction  of  cellular  tissue  in  disease  or  on  too  low  protein 
intake. 

Smetanka  (1911)  finds  that  any  protein,  even  if  purin-free, 
leads  to  increased  uric  acid  excretion,  a  fact  which  he  attributes  to 
changes  in  the  composition  of  the  cells  of  the  digestive  glands  dur- 
ing their  activity.  Ingestion  of  polysaccharides  causes  less  in- 
crease, corresponding  with  less  activity  of  digestion  called  for. 
Honey,  however,  causes  great  increase,  which  may  be  explained  by 
the  intense  activity  of  the  liver  cells  in  forming  glycogen.  Increase 
of  total  nitrogen  metabolism  does  not  in  itself  increase  uric  acid 
excretion. 

Schittenhelm  and  Seisser  (1911)  conducted  feeding  and  in- 
travenous injection  experiments  with  dogs  and  rabbits  showing  that 
sodium  thymonucleate  influences  but  slightly  the  outgo  of  uric  acid 
and  purin  bases,  while  it  prominently  increases  the  outgo  of  allan- 
toin ;  from  which  they  conclude  that  nucleic  acid  is  without  influence 
on  uric  acid  transformation,  especially  that  it  cannot  protect  uric 
acid  from  further  decomposition. 

In  the  observations  of  M.  Tschernoruzki  (1912)  with  dogs, 
both  nitrogen  and  phosphorus  excretion,  after  the  administration  of 
the  sodium  salt  of  nucleic  acid  (Merck's),  indicated  an  increased 
metabolism  of  phosphorized  body  tissues. 

The  behavior  of  iron-iodine-paranucleate  (iodtrif errin) ,  when 
fed  to  dogs  and  rabbits,  has  also  been  studied  by  Salkowski  (1913a). 

Hyperleucocytosis.  As  a  constant  accompaniment  of  the  tak- 
ing of  huclein-rich  food,  and  especially  of  the  injection  of  nuclein  or 


248  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

nucleic  acid,  and  closely  associated  with  increased  uric  acid  elimina- 
tion, is  an  increase  in  the  number  of  leucocytes  in  the  blood.  The 
high  phosphorus  output  at  the  time  of  such  nuclein  intake  has  been 
attributed  to  a  leucocyte  destruction  following  the  great  leucocyte 
formation.  Since  leucocytes  are  so  effective  in  battling  with  path- 
ogenic bacteria,  use  has  been  made  of  nuclein  injection  as  a  prophy- 
lactic measure  in  guarding  against  infection  in  preparation  for  sur- 
gical operations. 

In  1891  Horbaczewski  called  attention  to  this  accompaniment 
of  nuclein  feeding,  and  in  1892  summed  up  the  results  of  his  experi- 
ments thus  (we  quote  through  Schaumann) :  "The  only  observed 
symptom  of  nuclein  action  which  is  constant  in  normal  animals  and 
men  is  an  increase  t>f  leucocytes  in  the  blood,  which  may  reach  as 
high  as  100  percent.  This  leucocytosis  appears  soon  after  nuclein 
is  given,  decreases  gradually,  and  may  entirely  disappear  after  18-24 
hours.  The  nuclein  leucocytosis  appears  constant  also  in  different 
pathological  conditions." 

Schaumann  also  says  that,  "Goldscheider  and  Jacob  (Zeit.  f. 
klin.  Med.,  25,  1894)  had  somewhat  different  results  in  their  ex- 
periments on  these  phenomena.  They  used  carbolic-glycerin  ex- 
tracts of  different  organs,  which  they  applied  by  injection.  In  these 
cases  no  changes  in  the  blood  were  caused  by  the  extracts  from  pan- 
creas, liver,  kidneys,  or  thyroid  gland,  while  those  of  spleen,  bone 
marrow  and  thymus  gave  positive  effects,  though  differing.  The 
last  two  produced  a  pronounced  influence  in  the  direction  of  hypoleu- 
cocytosis,  while  the  spleen  extract  injection  caused  a  hyperleucocy- 
tosis,  the  intensity  and  duration  of  which  were  several  times  as 
great  as  with  the  giving  of  spleen  pulp  per  os." 

From  among  the  many  other  observations  on  leucocytosis  the 
following  may  be  of  interest.  Some  have  asserted  that  the  in- 
crease of  leucocytes  is  only  an  apparent  increase  due  to  a  transfer 
of  these  corpuscles  from  the  central  circulation  to  the  peripheral. 
Delano  Ames  and  A.  A.  Huntley  (1897)  disproved  this  theory  by 
their  experiments  on  dogs.  Comparison  was  also  made  of  the  re- 
spective numbers  of  the  three  varieties  of  leucocytes,  which  are  sup- 
posed to  be  three  stages  in  their  development — small  mononuclear, 
large  mononuclear,  and  polynuclear  cells.  It  was  noticed  that  the 
administration  of  ether  produced  an  increase,  the  extent  of  which 
seemed  to  depend  on  the  amount  of  ether  used.  "2.  Before  the 
administration  of  nuclein  the  counts  showed  that  the  number  of 
leucocytes  in  the  peripheral  and  in  the  central  circulation  was  prac- 
tically the  same 3.  Following  the  administration  of  the  nu- 
clein solution  there  was  immediately,  that  is  by  the  end  of  five  or 


PHOSPHORUS  METABOLISM  249 

ten  minutes,  a  noticeable  increase  in  the  number  of  leucocytes  in 
both  the  central  and  peripheral  circulation.  4.  At  this  time  the  per- 
centage increase  was  most  marked  in  the  young,  mononuclear  forms 
while  the  proportion  of  polynuclear  elements  was  proportion- 
ately low.  5.  The  longer  after  the  injection  of  nuclein  the  greater 
was  the  actual  increase  and  the  number  seemed  steadily  to  rise  in 
both  the  peripheral  and  central  circulation." 

According  to  Schittenhelm  and  Bendix  (1905),  yeast  nucleic 
acid,  and  a-thymus  nucleic  acid  injected  in  rabbits  as  the  sodium 
salt,  both  caused  at  once  hypoleucocytosis,  followed  in  6-8  hours  by 
great  hyperleucocytosis,  which  corresponds  with  Renner's  observa- 
tions (See  p.  258).,  According  to  Schaumann,  Walter  Hannes  ob- 
served as  high  as  144  percent  increase  of  leucocytes.  Renner's 
greatest  increase  was  even  higher,  452  percent. 

Meisen  (1909,  1911)  finds  that  nucleic  acid,  subcutaneously  in- 
jected in  growing  dogs,  produces  hyperleucocytosis,  without  injuri- 
ous accompaniments ;  that  the  relative  number  of  mononuclear  neu- 
trophils is  not  increased;  that  with  repeated  injections  there  is  an 
up-and-down  variation  in  the  number  of  red  blood  corpuscles  and  in 
the  haemoglobin  content;  and  that  injections  kept  up  for  a  long 
time  cause  a  greater  firmness  in  growing  bones. 

Kaupp  (1911)  reports  certain  blood  observations  on  horses  af- 
ter nuclein  injections,  the  conclusions  from  which  may  be  summed 
up  thus :  Nuclein  increases  the  number  of  leucocytes  in  the  blood,  the 
number  of  polymorphonuclear  leucocytes,  and  the  power  of  phagocy- 
tosis of  the  individual  leucocyte.  Nuclein  decreases  the  time  of  co- 
agulation of  the  blood.  To  obtain  the  best  results  in  cases  of  infec- 
tious diseases,  nuclein  should  be  given  hypodermically.  To  obtain 
desirable  results  in  cases  of  hemorrhage,  the  nuclein  should  be  given 
intravenously. 

M.  Tschernoruzki  (1912)  made  a  special  study  of  the  effects  of 
introduction  of  the  sodium  salt  of  nucleic  acid  in  dogs.  The  dogs 
receiving  the  nucleate  either  per  os,  subcutaneously,  intraperitoneal- 
ly  or  intravenously  showed  a  more  or  less  marked  hyperleucocytosis, 
and  a  somewhat  larger  number  of  red  corpuscles  than  an  untreated 
dog.  As  judged  from  the  number  of  leucocytes  found,  the  strong- 
est hyperleucocytosis  resulted  in  the  case  of  intravenous  injection 
(353.3  percent  of  the  value  in  the  control  dog),  next  in  the  case  of 
subcutaneous  administration  (204.8  percent),  then  intraperitoneal 
introduction  (187.6  percent)  and  last,  in  the  case  of  introduction  per 
os  (162.5  percent).  The  animals  quickly  acquired  a  certain  immun- 
ity to  considerable  quantities  of  the  nucleate. 


250  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Timoshok  (1912)  studied  the  influence  of  sodium  nucleinate 
(from  yeast)  on  rabbits  infected  with  Staphylococcus  pyog.  aur.  The 
nucleinate  caused  a  rapid  polynuclear  hyperleucocytosis,  and  altera- 
tion of  enzyme  functions,  the  latter  varying  either  directly  or  in- 
versely as  the  changes  occurring  in  Staphylococcus  infection  without 
sodium  nucleinate.  There  were  noted:  (1)  Parallel  increase  in  lipoly- 
tic and  catalytic  activity  of  the  organs;  (2)  parallel  decrease  in  the 
sugar-synthesizing  energy  of  the  organs,  and  the  diastatic  and  cata- 
lytic energy  of  the  blood ;  (3)  increase  in  the  lipolytic  power  of  the 
serum  (a  decrease  occurred  in  the  absence  of  sodium  nucleinate) ; 
(4)  decrease  in  the  diastatic  and  sugar-synthesizing  energy  of  the 
organs,  and  the  anti-trypsin  content  of  the  serum  (an  increase  oc- 
curred when  sodium  nucleinate  was  absent). 

MISCELLANEOUS  EFFECTS  OF  NUCLEAR  MATERIAL 

.  Milroy  and  Malcolm  (1899)  reported  a  histological  study  with 
rabbits  and  guinea  pigs  which  was  concerned  with  the  intracellular 
wanderings  of  granulated  leucocytes  under  the  influence  of  nucleic 
acid,  thymic  acid,  adenin,  guanin,  cytosin  and  metaphosphoric  acid. 

Bang  (1901b)  tried  the  effects  of  injection  of  guanylic  acid  and 
of  /?-nucleoprotein  with  dogs.  Both  of  these  compounds  greatly 
reduced  the  rate  of  coagulation  of  blood,  the  nucleoprotein  most 
markedly.  Blood  pressure  fell  off  noticeably  after  the  injection. 
Immediately  after  the  injections  the  dogs  were  always  restless,  and 
breathed  irregularly  and  quickly;  then  the  excitement  passed  and 
was  followed  by  partial  narcosis  for  some  minutes.  In  the  case  x>f 
guanylic  acid  injection,  the  urine  became  alkaline  about  half  an  hour 
after  injection.  After  24-48  hours  it  was  again  acid.  There  was 
no  more  ammonia  present  than  normal,  and  the  alkaline  reaction 
could  not  be  explained.  Albumin  was  present  to  the  extent  of  1-3 
parts  per  thousand,  which,  it  was  thought,  might  be  due  to  the  low- 
ering of  blood  pressure  or  to  the  acid  having  toxic  action  on  the 
kidneys. 

Galeotti  (1900)  reports  tests  of  the  effects  of  bringing  nucleo- 
proteins  extracted  from  animal  tissues  or  from  bacteria,  into  con- 
tact with  different  animal  organs  or  self-moving  cells.  The  report 
is  that  nucleoproteins  exercise  considerable  influence  on  protoplasm 
in  general,  the  results  of  the  influence  varying  according  to  the  cells 
concerned.  For  some  cells  the  influence  was  of  the  nature  of  ex- 
citement, and  for  some  it  was  paralyzing. 

Mendel,  Underhill  and  White  (1903)  report  experimental 
studies  of  the  physiological  effects  of  nucleic  acid,  particularly  that 
from  wheat  embryo,  tritico-nucleic  acid.  Observations  of  the  fol- 
lowing kinds  are  reported : 


PHOSPHORUS  METABOLISM  251 

(1)  Manometer  tracings  of  blood  pressure  after  intravenous 
injections,  (2)  "coagulation  time  of  blood  taken  from  femoral  vein 
after  injections,  (3)  coagulability,  composition  and  rate  of  flow  of 
lymph  from  thoracic  duct,  (4)  immunity  effects,  (5)  urine  examina- 
tion after  feeding,  and  intravenous,  intraperitoneal,  subcutaneous 
and  rectal  injection,  (6)  urine  and  feces  examination  and  leucocyte 
count  after  feeding  and  rectal  injection. 
The  authors'  concluding  summary  is: 

"The  more  important  observations  recorded  in  this  paper  indi- 
cate that  the  vegetable  nucleic  acid  obtained  from  the  wheat  embryo 
resembles,  in  its  physiological  effects,  the  guanylic  acid  of  the  pan- 
creas. Introduced  in  sufficient  doses  into  the  circulation,  it  may 
produce  a  fall  in  arterial  pressure ;  a  change  in  the  coagulability  of 
the  blood ;  an  increase  in  the  flow  of  lymph  and  a  change  in  its  com- 
position; and  perhaps,  also,  a  degree  of  immunity  toward  subse- 
quent injections. 

"The  ingestion  of  nucleic  acid  is  followed  in  man  by  an  in- 
creased output  of  uric  acid,  and  in  the  dog  by  the  excretion  of  al- 
lantoic These  products  correspond  in  either  case  to  only  a  portion 
of  the  purin  radicals  introduced.  In  animals  allantoin  excretion 
was  also  observed  after  the  introduction  of  vegetable  nucleic  acids 
into  the  body  per  rectum,  intravenously,  intraperitoneally,  and  sub- 
cutaneously.  Some  features  of  intermediary  purin-metabolism  are 
discussed." 

Guerrini  (1903)  made  injections  of  1  percent  aqueous  or  sodium 
carbonate  solutions  of  nucleoproteins  from  the  liver  and  brain  of 
white  mice  and  of  dogs  into  the  peritoneal  cavity  of  dogs  and  white 
mice,  following  with  histological  examination  of  the  organs  of  the 
animals  after  different  intervals  of  time,  varying  from  one-half 
hour  to  72  hours.  Both  excitations  and  degenerations  were  always 
found  to  have  taken  place  in  the  parenchymatous  cells  of  the  livers 
and  of  the  kidneys,  the  intensity  and  the  time  of  appearance 
of  the  changes  being  in  evident  relation  to  the  quantity  of  nucleopro- 
tein  injected.  Both  kinds  of  changes  were  evident  in  both  the 
nuclei  and  the  cytoplasm  of  the  affected  cells. 

Enea  (1903)  carried  out  injection  experiments  with  rabbits, 
from  which  he  concluded  that  nuclein  extracted  from  pathogenic 
and  non-pathogenic  bacteria  has  a  toxic  action;  that  in  maximum 
doses  it  causes  immediate  death  by  destruction  of  leucocytes  and  co- 
agulation of  the  blood,  while  in  lesser  doses  it  produces  gradual  pro- 
gressive destruction  of  leucocytes,  but  not  general  coagulation, 
death  occurring  after  different  lengths  of  time  that  are  proportion- 
al to  the  quantities  of  nuclein  injected.  It  also  increases  the  nor- 
mal bactericidal  power  of  blood  serum,  the  effect  not  being  specific 
for  the  organism  from  which  the  nuclein  was  taken. 


252  .  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Schittenhelm  and  Bendix  (1904)  found  gelatinizing,  unab- 
solved nucleic  acid  in  the  urine  of  rabbits  after  the  subcutaneous 
injection  of  the  sodium  salt  of  a-thymonucleic  acid;  and  the  same 
acid  injected  in  the  ear  vein  of  rabbits  led  to  the  appearance  t)f  red 
and  white  corpuscles,  nucleic  acid  cylinders,  and  the  unchanged  nu- 
cleate in  the  urine ;  also  purin  bases  were  present ;  the  kidneys  were 
disturbed.  Addition  of  uric  acid  to  the  nucleate  injected  made  lit- 
tle difference,  save  that  uric  acid  was  included  in  the  nucleic  acid 
cylinders. 

Desgrez  and  Zaky  (1904a,  1904b,  1904c,  1905)  concluded  that 
in  guinea  pigs  and  dogs  the  nuclein  of  yeast,  or  its  nucleic  acid,  fa- 
vored retention,  especially  of  proteins,  and  a  mineralization  of  the 
skeleton.  (See  Growth' and  Comp.  of  An.  as  Aff.  by  Comp.  of  Glyc.) 

Snowman  (1905)  mentions  an  experiment  of  Miyake,  reported 
in  Med.  Annual  for  1905,  in  which  the  injection  of  1  c.c.  of  0.5  per- 
cent solution  of  nucleic  acid  into  the  peritoneum  of  a  guinea  pig 
caused  the  animal  to  resist  a  subsequent  inoculation  with  Bacillus 
coli  of  20  times  the  normal  minimum  fatal  dose. 

Pouchet  and  Chevalier  (1906)  report  that  injection  of  nucleic 
acid  (from  pancreas  and  from  fish  roe)  causes  an  increase  of  leuco- 
cytes. An  intravenous  injection  of  0.004  gm.  per  kilogram  body 
weight  in  dogs  causes  acceleration  of  heart  beat.  With  a  larger  dose 
(0.020  gm.  per  kilo.)  an  irritating  action  is  produced  upon  the  endo- 
cardium, and  there  is  a  decrease  in  blood  pressure,  and  a  slower 
rate  of  heart  beat.  After  a  certain  time  the  original  blood  pressure 
is  regained,  and  the  heart  beat  becomes  normal.  The  effects  are 
thought  to  be  due  to  action  on  the  central  nervous  system. 

Knapp  (1908)  states,  after  his  experiments  with  rats,  that  con- 
tinued use  of  sodium  nucleate  causes  nephritis  and  fatty  degenera- 
tion of  the  liver. 

Achard  and  Redfield  (1911)  used  a  nuclein  solution  as  a  wound 
dressing  and  consider  that  it  promoted  healing.  Much  literature 
that  we  have  not  seen,  on  the  bactericidal  effect  of  nucleins,  is  re- 
viewed. 

M.  Tschernoruzki  (1911,  1912,  also  Czernoruzky,  1912)  has  re- 
cently reported  an  interesting  study  on  the  action  of  nucleic  acids 
on  the  fermentative  processes  of  the  animal  organism.  Think- 
ing that  the  resistance  which  the  organism  acquires  against  infec- 
tion from  the  use  of  sodium  salts  of  nucleic  acids  is  not  due  to  leu- 
cocytosis  alone,  because  it  differs  from  the  effect  of  aleuronate, 
which  produces  equally  great  leucocytosis,  but  may  be  in  part  due  to 
alterations  of  the  fermentative  functions  of  the  organs,  he  put  to 
test  the  individual  organs  of  dogs,  after  long  periods  of  ingestion  or 
injection  of  nucleic  acid. 


PHOSPHORUS  METABOLISM  253 

Five  young  poodles  of  the  same  litter  were  used.  One  was  kept 
as  a  control,  and  the  other  four  received  Merck's  sodium  nucleate, 
from  yeast :  No.  1  intravenously,  No.  2  intraperitoneal^,  No.  3  sub- 
cutaneously,  and  No.  4  by  mouth.  Treatment  was  begun  at  about 
the  second  month  and  continued  4*4-5  */£  months,  with  increasing 
doses  up  to  1.5  gm.  per  kilogram  of  body  weight.  Rise  of  temper- 
ature and  leucocytosis,  as  results  of  the  treatments,  were  noted.  All 
of  the  dogs  were  killed  and  the  organs  removed,  ground,  dried,  ex- 
tracted with  physiological  salt  solution,  and  the  extract  tested  for 
ferments.  The  organs  examined  were;  liver,  lungs,  spleen,  brain, 
kidneys,  muscles  and  thymus ;  and  the  following  were  the  tests  ap- 
plied : 

Fermeats  tested  for  Tests 

Protease    (Tryptase)  Amount   of  extract  required   to   digest   completely   0.5   c.c. 

casein  solution 

Amylase  Amount  of  extract  required  for  digestion  of  starch  to  dis- 

appearance of  iodine  reaction 

Diastase  (Dextrinase)  Amount  of  sugar  produced  from  starch 

Oatalase  Potassium  permanganate  method   (Results  are  not  reported 

because  conditions  of  preparation  of  organ  extract 
would  vitiate  results) 

Nuclease  Inorganic    phosphorus    produced    from   nucleic    acid    in    48 

hrs.  at  37.5°C. 

Lipase  "  Acid  split  off  from  1  percent  monobutyrate  solution 

Jjecithase  Acid   split    from    2    percent   lecithin   emulsion    (All   results 

here  were  negative.) 

The  author's  conclusions  are  in  part  as  follows : 
Introduction  of  nucleic  acid  into  the  animal  organism  has  a 
marked  influence  on  its  fermentative  activities.  As  to  the  signifi- 
cance of  the  manner  of  its  introduction — the  least  effect  on  ferment- 
ative functions  was  observed  with  subcutaneous  and  the  greatest 
with  intravenous  injection.  As  to  the  ferment — the  greatest 
changes  were  found  with  the  amylolytic  ferment.  With  regard 
to  the  organs — the  greatest  variations  in  the  sense  of  increase  of 
fermentative  energy  were  in  the  brain,  the  lungs,  the  muscles  and 
the  thymus.  In  the  brain  the  value  for  amylase  was  400  times  as 
high  as  the  normal,  for  diastase  4.4  times,  and  for  protease  10  times. 
In  the  lungs  the  value  for  amylase  was  increased  250  times,  in  the 
muscles — for  amylase  6.4  times,  in  thymus — for  lipase  2.5  times. 
The  brain  considerably  exceeded  the  other  organs  in  the  number  of 
ferments  evident,  as  well  as  in  the  amount  of  change  in  their  action. 
Merck's  sodium  salt  of  yeast  nucleic  acid  produced  no  evil  effects  in 
the  animal  organism  under  continual  use  of  large  doses,  and  with 
different  ways  of  introduction.  Perhaps  the  therapeutic  signifi- 
cance of  nucleic  acid  preparations  in  various  diseases  finds  some  ex- 
planation in  the  facts  here  reported.  Especially  in  progressive 
paralysis,  the  benefit  may  be  due  to  changes  in  the  fermentative 
functions  of  the  brain.  The  strengthening  of  the  neucleolytic 
function  in  the  blood  was  greatest  in  the  polynuclear  white  corpus- 
cles.     Of  the  organs  examined,  there  seemed  to  be  great  increase 


254  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

of  nuclease  content  in  the  thymus,  pancreas  and  liver,  less  of  an  in- 
crease in  bone-marrow  and  spleen,  and  decrease,  or  but  slight  in- 
crease, in  brain,  lungs,  kidneys  and  muscles.  The  dog  which  re- 
ceived the  nucleate  by  mouth  showed  the  greatest  nuclease  varia- 
tion from  normal,  considering  all  the  organs  together;  and  the  one 
receiving  it  subcutaneously,  the  least. 

In  addition  to  these  effects  on  ferments,  and  to  a  hyperleucocy- 
tosis  and  increased  purin  metabolism,  Tschernoruzki  (1912)  noted 
in  the  dogs  a  rise  in  body  temperature  and  certain  derangements  in 
general  condition,  whatever  the  method  of  introduction.  In  the 
case  of  parenteral  injection,  doses,  which  approached  the  value  of  1 
gm.  per  kg.  of  body  weight  produced  rather  marked  disturbances, 
such  as  loss  of  appetite,  apathy,  etc.  After  a  dose  of  1.5  gm.  per 
kg.  of  body  weight  the  intravenously  injected  dog  showed  symptoms 
like  those  of  poisoning,  but  the  dog  receiving  it  by  way  of  mouth 
showed  no  disturbance  of  general  condition,  with  the  exception  of 
diarrhoea. 

At  the  close  of  this  second  paper  Tschernoruzki 's  conclusions 
were  as  follows:  There  is  a  strong  analogy  in  the  action  upon  the 
organism  of  nucleic  acid,  and  infection.  Both  result  in  hyperleuco- 
cytosis,  rise  in  temperature,  disturbance  of  general  condition,  and 
increase  of  metabolism;  and  both  react  upon  the  fermentative 
processes,  and  bring  about  a  certain  degree  of  immunity.  Nucleic 
acid  is  then  apparently  an  agent  which  stimulates  the  natural  pro- 
tective agencies  of  the  organism,  especially  the  leucocyte  function. 
This  explains  the  therapeutic  significance  of  nucleic  acid  and  gives 
basis  for  its  use  in  medicine.  His  final  conclusion  is  that  "by  intro- 
duction of  nucleic  acid  into  the  animal  organism  the  nucleolytic 
function  of  the  latter  is  increased." 

Brossa  (1912)  made  an  attempt  to  determine  what  he  calls  the 
biological  value  of  a-nucleic  acid  of  thymus,  for  a  dog  and  for  hens, 
by  applying  to  the  nitrogen  intake  and  output  the  following  form- 
ula, which  he  attributes  to  Thomas : 

d_[C-(a-b)] 

100 

a-b 

A  nitrogen-free  diet  was  fed  for  a  brief  time,  followed  by  a  like 
diet  with  sodium  nucleate  added,  and  to  the  data  obtained  the  form- 
ula was  applied  by  letting  "a"  equal  the  nitrogen  taken  in  the  food, 
"b"  that  excreted  in  the  feces,  "c"  that  in  the  urine  on  the  nitrogen- 
containing  food,  and  "d"  that  in  the  urine  on  the  nitrogen-free  diet. 
In  this  way  it  is  calculated  that  the  biological  value  of  the  nucleic 
acid  for  the  dog  is  60  percent,  and  for  the  hens  80  percent. 


PHOSPHORUS  METABOLISM  255 

Goubau  and  VanGoethem  (1913)  determined  by  intravenous  in- 
jection experiments  with  dogs  that  the  effects,  on  the  circulation, 
of  injecting  nuclein,  and  nucleohistone  are  quite  comparable  with 
those  of  proteins  in  general.  The  primary  effect  is  thromboplastic, 
and  is  followed  by  a  more  lasting  antithrombic  influence.  Nucleins 
paralyze,  slightly,  the  motor  center  of  the  heart  immediately  after 
the  injection,  with  an  enormous  increase  in  the  frequency  of  the 
beats,  followed  by  a  secondary  enfeebling  of  the  beats. 

See  also  Bokay  (1877),  Gumlich  (1894),  Jacob  and  Bergell 
(1898),  A.  Neumann  (1898). 

SIGNIFICANCE  OF  NUCLEOPROTEINS  AND  NUCLEIC  ACIDS  AS  AC- 
TIVE AGENTS  IN  SPECIFIC  BODY  PROCESSES 

From  the  mere  fact  of  the  presence  of  nucleoproteins  at  the 
heart  of  every  cell,  it  may  be  inferred  that  they  are  of  great  import- 
ance in  the  life  and  the  activity  of  cells,  as  Kossel  has  repeatedly  re- 
marked. As  to  just  what  is  their  part  we  cannot  say,  but  we  may 
quote  Gustav  Mann  as  to  one  probability.  As  a  result  of  his  ex- 
tensive study  and  observation,  chemical  and  microscopical,  he  says 
(Chemistry  of  Proteids,  1906,  p.  454) : 

"What  we  call  life  is  simply  the  manifestation  of  special  chem- 
ical compounds,  and  if  we  see  microscopically  that  every  manifest- 
ation of  metabolism  is  accompanied  by  enormous  changes  in  the  nu- 
cleoproteids, and  that  the  rapidity  with  which  nucleoproteids  or  the 
nuclear  basophil  chromatin  reacts  to  food  substances  is  directly  pro- 
portional to  the  ease  with  which  the  food  is  absorbed,  we  cannot  ar- 
rive at  any  other  conclusion  but  that  the  nucleoproteids  are  the 
agencies  by  which  amino-acids  are  built  up  into  the  cell  plasm." 

Herlitzka  and  Borrino  (1902a,  1902b,  1903),  following  Bottazzi, 
attribute  catalytic  actions  to  the  nucleohistone  of  cell  nuclei  and  the 
nucleoprotein  of  the  cytoplasm.  They  tested  the  isolated  nucleo- 
histones  and  nucleoproteins  of  liver,  thymus  and  kidney  as  to  their 
power  to  decompose  carbonates,  and  to  free  the  C02  (the  power  was 
possessed  by  both),  to  break  down  haemoglobin  (possessed  by  both 
kinds  of  compounds,  but  only  those  of  liver),  to  form  urea 
(possessed  by  neither),  and  to  digest  sugars  (most  action  by  com- 
pounds from  kidney,  especially  from  its  nuclei). 

Fibrin  ferment  and  blood  coagulation  action  have  been  ascribed 
to  nucleoproteins  and  nucleohistones  of  blood  and  of  muscle.  The  in- 
jection of  nuclear  matter  from  various  sources  has  been  followed 
by  intravascular  coagulation.  (See  Halliburton,  1892;  Halliburton 
and  Brodie,  1894 ;  Pekelharing,  1895,  1896 ;  Lilienf eld,  1895 ;  Huis- 
kamp,  1901a).  Other  substances,  however,  introduced  in  this  way 


256  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

have  also  led  to  such  consequences.  It  is  Pekelharing's  (1895,1896, 
1914)  idea  that  the  fibrin-ferment,  thrombin,  forms  from  a  nucleo- 
protein  of  the  blood  or  organ,  it  being  perhaps  a  calcium  compound 
of  the  nucleoprotein. 

On  the  other  hand,  several  observations  which  have  been  quoted 
above  show  that  injections  of  nucleic  acids  affect  the  blood  in  such 
manner  that  its  coagulation  is  considerably  retarded.  Doyon  and 
his  associates  (Doyon,  1912 ;  Doyon  and  Dubrulle,  1912 ;  Doyon  and 
Sarvonat,  1912a,  1912b,  1912c,  1913a,  1913b,  1913c,  1913d,  1913e, 
1913f ;  Doyon,  Dubrulle  and  Sarvonat,  1912,  and  others)  have  re- 
ported investigations  of  this  power  in  vitro.  Anticoagulating 
bodies  were  extracted  from  liver,  testicles,  intestine,  thymus,  pan- 
creas, mesenteric  ganglia,  beer-yeast,  blood  corpuscles  of  birds,  and 
haematogen  of  egg.  Most  of  these  were  nucleic  acids  or  nucleo- 
proteins ;  and  it  is  held  that  a  phosphorized  nucleus  is  essential  to 
the  anticoagulating  power.  The  degree  of  effect  of  nucleic  acid  was 
compared  (Doyon  and  Sarvonat,  1913c)  with  that  of  ortho-,  pyro- 
and  meta-phosphoric  acids,  with  the  resulting  observation  that  or- 
thophosphate  does  not,  under  the  conditions  used,  exercise  any  sen- 
sible anticoagulating  action,  while  the  pyro-  and  meta-phosphates  do. 
The  amounts  of  sodium  metaphosphate  and  sodium  nucleate  with 
the  same  content  of  phosphorus  showed  the  same  degree  of  antico- 
agulating action,  and  these  observations  are  taken  as  supporting  the 
view  that  the  phosphorus  of  nucleic  acids  is  present  as  metaphos- 
phoric  acid.  Glycerophosphoric  acid  and  lecithin  were  inactive  in 
corresponding  amounts. 

Pepsin  had  been  considered  by  some  to  be  a  nucleoprotein,  but 
Pekelharing  (1902)  showed  that  the  phosphorus  content  of  the 
preparation  may  be  reduced  by  purification  processes,  and  that  in  all 
probability  phosphorus  is>  not  a  constituent  of  pure  pepsin.  Giacosa 
and  Dezani  (1909)  also  report  the  isolation  from  pig  stomach  of  a 
digestive  enzyme  which  is  still  active  when  free  from  phosphorus. 

THERAPEUTIC  AND  PROPHYLACTIC  USE  OF  NUCLEINS  AND 

NUCLEIC  ACIDS 

Some  of  the  effects  of  isolated  nucleic  acids  in  the  diet  are  bene- 
ficial and  some  otherwise.  If  the  body  be  in  normal  condition,  they 
are  well  used  and  are  favorable  to  nitrogen  and  phosphorus  reten- 
tion; and  in  pathological  cases  the  same  is  usually,  but  not  always, 
true.  When  the  amount  of  uric  acid  in  the  system  is  excessive, 
nucleins  should  be  avoided.  In  special  cases  they  may  be  of  de- 
cided advantage,  especially  if  injected,  because  of  their  effects  upon 
the  circulation  and  upon  the  number  of  blood  corpuscles,  particularly 
the  leucocytes,  and  upon  the  power  of  phagocytosis.      They  have 


PHOSPHOKUS  METABOLISM  257 

been  used  with  apparent  benefit  in  several  diseases,  and  most  strik- 
ingly as  a  prophylactic  measure  before  surgical  operations  and 
child-birth.  See  the  reviews  of  Aulde  (1900),  Martinet  (1902b)  and 
Meisen  (1911)  of  the  literature  of  nucleins  and  nucleic  acids  from 
the  point  of  view  of  therapeutics. 

Larned  (1902)  describes  a  series  of  iron,  copper,  mercury  and 
silver  nucleids  as  reconstructives  and  tonics,  and  as  possessed  with 
the  power  to  increase  physiologic  resistance,  and  functional  activity 
of  secretory  organs;  and  Burnet  (1903)  says  that  the  metals  of 
these  salts  are  better  used  when  administered  in  this  form  than  in 
other  forms. 

Tomlinson  (1897)  reports  that  in  infantile  scrofula,  and  other 
cases  of  enlarged  lymphatic  glands,  with  discharges,  the  stimulation 
of  leucocytosis  prevents  suppuration,  if  it  has  not  begun,  and  in- 
creases it,  if  it  has  begun,  so  that  a  more  complete  cure  is  brought 
about. 

Laumonier  (1905)  also  reports  as  being  brought  nearer  to  nor- 
mal, under  his  observation,  certain  more  or  less  definitely  estab- 
lished physiological  states. 

Backus  (1907)  reports  apparent  benefit  in  the  cure  of  mange. 

Joseph  Hoppe  (1907a)  made  artificial  nucleic  acid  preparations 
from  yeast  cells  for  use  with  children  with  nervous  disease. 

Donath  (1909)  discusses  the  benefits  from  nuclein  injection  in 
the  treatment  of  progressive  general  paralysis.  He  states  that  the 
injection  at  intervals  of  5-7  days  of  50-100  c.c.  of  a  2  percent  solu- 
tion of  sodium  nucleate,  with  an  equal  quantity  of  sodium  chloride 
added,  is  effective  in  the  initial  stages  of  this  disease.  The  injec- 
tions caused  a  rise  of  temperature  and  an  increase  of  leucocytes, 
with  increased  oxidation,  which  brought  about  a  destruction  of  the 
poisonous  metabolic  products  formed  by  the  disease. 

Bachrach  and  Bartel  (1907)  conducted  experiments  in  which 
yeast  nucleic  acid,  added  to  protein  media  containing  tubercle  ba- 
cilli (from  man),  increased  the  speed  of  reduction  of  their  virulence. 
On  the  other  hand,  the  addition  of  yeast  nucleic  acid  to  suspensions 
of  the  bacilli  in  distilled  water  helps  to  retain  the  virulence  of  the 
organisms,  which  in  distilled  water  alone  become  avirulent  in  but  a 
few  days. 

Ward  (1910)  reported  favorable  results  from  intravenous  in- 
jection of  sodium  tritico-nucleate  in  physiological  salt  solution  in  the 
treatment  of  tuberculosis.  The  anaemia  and  oxygen-starvation  of 
the  tuberculous  patients  are  associated  with  a  low  specific  gravity 
of  the  blood,  due  to  a  decrease  in  the  number  of  red  corpuscles,  and, 
at  the  same  time,  the  appearance  of  increasing  numbers  of  poikilo- 


258  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

cytes.  It  is  said  that  under  the  influence  of  the  nuclein-saline  so- 
lution the  poikilocytes  rapidly  disappear  from  the  circulation, 
the  haemoglobin  being  eliminated  by  the  liver  as  bili- 
rubin and  biliverdin;  the  deformed  cells  being  replaced 
by  new  and  healthy  erythrocytes  more  rapidly  than  is 
possible  under  any  other  form  of  treatment  as  yet 
devised.  This  author,  then,  thinks  that  the  administration  of  nu- 
clein  produces  its  good  effects,  not  chiefly  by  inducing  leucocytosis, 
but  by  effecting  an  upbuilding  of  erythrocytes  and  haemoglobin. 

Though  nucleins  and  nucleic  acids  may  have  bactericidal  action 
in  themselves  (see  Vaughan,  Novy  and  McClintock,  1893;  Kossel, 
H.,  1894),  their  use  to  prevent  infection,  in  cases  of  exposure  of 
the  tissues  to  pathogenic  bacteria,  is  probably  to  be  attributed  more 
largely  to  their  leucocytosis-producing  power.  Rentier  (1905)  re- 
ported such  use  of  yeast  nucleic  acid  in  133  surgical  cases.  Usually 
50  c.c.  of  a  2-percent  solution  of  the  acid  in  sodium  chloride  solution 
was  injected  in  the  breast  within  24  hours  before  the  operation. 
There  was  a  brief  hypoleucocytosis,  followed  by  a  pronounced  hyper- 
leucocytosis.  Apparently  the  resistance  against  Bacterium  coli  and 
other  pathogenic  bacteria  was  increased.  The  increase  of  leucocyte 
count  amounted  to  from  9  to  452  percent  (with  the  exception  of  S1/^ 
percent  in  one  case  where  the  initial  count  was  very  high) ;  in  4  cases 
there  was  a  lowering  of  leucocyte  count,  at  least  temporary,  of  9-14 
percent;  the  average  increase  in  121  cases  was  118  percent.  For 
certain  undesirable  effects   to  be  guarded  against  see  the  original. 

See  also  Pollak  (1906) ;  Hannes  (1906) ;  Anzilotti  (1911) ; 
Achard  and  Redfield  (1911).;  also  (through  Schaumann)  v.  Mikulicz 
(Archiv  f.  klin.  Chirurgie  73,  Nr.  2),  Miyake  (Mitteil,  aus  den 
Grenzgebieten  der  Medizin  und  Chirurgie  13,  Nr.  14,  15)  and 
Dudgeon  and  Ross  ( Amer.  Jour,  of  the  Med.  Sciences  1906,  17 ; 
Presse  medicale  1906,  569). 

SUMMARY 

Very  little  is  known  as  to  the  details  of  the  anabolic  processes 
in  the  body ;  but  it  seems  probable,  though  not  proved,  that  the  ani- 
mal organism  is  capable  of  synthetic  formation  of  nucleins  and  nu- 
cleoproteins,  even  without  purins  being  supplied  as  such.  Obser- 
vations reported  as  evidence  of  such  apparent  synthesis  with  inad- 
equate supply  of  purins  are  of  the  following  types :  The  development 
of  the  genitalia  of  fish  while  no  food  is  taken ;  the  growth  of  young 
mammalia  when  milk  is  the  only  article  of  diet ;  the  transformations 
within  eggs  and  embryos ;  and  apparently  normal  growth  on  experi- 
mental purin-free  (or  low-purin)  diet.  However,  the  evidence  is 
not  complete  on  any  one  of  these  points. 


PHOSPHORUS  METABOLISM  259 

As  to  katabolic  processes  acting  on  nucleoproteins,  nucleins  and 
nucleic  acids,  tests  in  vitro  by  isolated  enzymes  or  by  fluids  taken 
from  the  digestive  tract  indicate  that,  though  proteins  may  be  sep- 
arated from  nucleoproteins,  and  nucleins  or  nucleic  acids  may  be 
dissolved  by  gastric  juice,  this  juice  does  not  cause  cleavage;  that 
by  activated  pancreatic  juice  (trypsin)  they  are  more  readily  dis- 
solved, which  solution  may  be  due  to  transformation  of  the  a-form 
to  the  /?-form,  or  the  breaking  up  of  polynucleotides  into  mononu- 
cleotides, though  there  may  be  slow  cleavage  of  the  nucleic  acids, 
but  not  a  complete  breaking  down  such  as  frees  the  reducing  sub- 
stance and  the  purin  bases;  that  intestinal  juice  cleaves  more  rap- 
idly, and  frees  phosphoric  acid,  but  does  not  break  down  the  gluco- 
side  complex  of  carbohydrate  and  base;  that  within  the  intestinal 
wall  cleavage  may  be  carried  to  the  state  of  free  purins ;  and  that 
the  intestinal  bacteria  may  not  only  bring  about  complete  cleavage, 
but  may  further  oxidize  and  transform  the  purins. 

Tests  in  vitro  with  blood  and  organ  extracts  indicate  that  cleav- 
age may  take  place  in  the  plasma  of  practically  all  organs  such  that 
the  phosphoric  acid  and  the  purins  are  freed  from  the  carbohydrate, 
and  that  the  purins  may  then  be  further  altered,  though  the  extent 
of  such  alteration  differs  with  different  organs,  with  different 
species,  and  probably  also  with  different  nucleic  acids. 

Animal  experimentation,  giving  fairly  direct  evidence  as  to 
what  actually  occurs  in  the  particular  animal  under  the  particular 
conditions  of  observation,  also  indicates  that,  in  the  stomach,  nu- 
cleoproteins and  nucleic  acids  are  not  broken  down,  but,  in  large 
part,  go  into  solution  as  such ;  that  in  the  intestine  there  is  some 
breaking  down;  that  this  breaking  down  cleaves  off  phosphoric  acid 
from  the  carbohydrate-base  complex,  but  does  not  free  the  purins ; 
and  that  nucleic  acids  and  their  cleavage  products  are  absorbed 
chiefly  from  the  lower  portions  of  the  small  intestine  (the  lower 
jejunum  and  the  ileum). 

We  may  suppose,  then,  that  nucleins  or  nucleic  acids  are  in  con- 
siderable part  taken  up  by  the  walls  of  the  digestive  tract  without 
change,  and  in  part  also  after  partial  cleavage,  and  that  cleavage 
may  be  carried  further  within  the  intestinal  walls,  or  at  the  place 
of  storage. 

The  ingestion  or  injection  of  nuclein  or  nucleic  acids  is  fol- 
lowed by  an  increased  purin  output  from  the  body,  which  probably 
originates  in  part  in  the  purin  component  of  the  nucleic  acid  intro- 
duced, and  in  part  in  the  nuclear  matter  of  cells  which  are  broken 
down  as  a  result  of  the  introduction  of  this  substance.  The  diges- 
tion itself  may  call  for  cell  destruction  which  produces  this  nuclear 


260  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

matter,  or  the  increased  formation  of  leucocytes  may  be  accompan- 
ied by  an  increased  destruction  of  such  cells.  There  is  also  an  in- 
creased phosphate  elimination. 

Another  constant  accompaniment  of  the  taking  of  nuclein-rich 
food  is  a  marked  increase  in  the  number  of  leucocytes,  in  the  blood, 
following  a  brief  period  of  decrease.  The  effect  is  more  pronounced 
when  the  nucleic  acid  is  injected.  It  takes  place  soon  after  the  in- 
troduction and  lasts  for  several  hours,  affecting  all  the  forms  of  leu- 
cocytes. The  phagocytic  power  of  the  individual  leucocytes  is  also 
said  to  be  raised.  Advantage  is  taken  of  this  favorable  result  in 
the  injection  of  nuclear  matter  as  an  aid  in  combating  infection. 

The  blood  shows  other  effects  of  nucleic  acid  injection  which 
are  more  or  less  temporary,  and  more  or  less  variable  with  the  con- 
ditions of  operation,  and  with  the  amount  and  kind  of  acid  used. 
There  may  be  an  increase  in  the  red  corpuscles,  a  fall  in  blood  pres- 
sure, and  a  change  in  coagulability.  While  nuclein  injection  is  said 
to  increase  the  power  to  resist  bacteria,  it  seems  to  be  somewhat 
toxic  itself.  One  author  (Tschernoruzki)  has  noted  favorable  ef- 
fects upon  the  fermentative  powers  (amylolytic,  diastatic,  catalytic, 
nucleolytic  and  lipolytic)  of  the  organs  as  a  result  of  the  introduc- 
tion of  nucleic  acid,  either  intravenously,  intraperitoneally,  subcu- 
taneously,  or  by  way  of  the  mouth.  This  effect  may  explain  some 
of  the  favorable  results  of  the  therapeutic  use  of  nucleins  and  nu- 
cleic acids. 

The  specific  activities  of  nucleoproteins  in  body  cells  are  not  de- 
termined; but  as  they  are  so  universally  present  as  the  main  con- 
stituents at  the  points  of  most  pronounced  manifestation  of  life — 
the  cell  nuclei — it  cannot  be  doubted  that  they  are  vital,  active 
agents. 

METABOLISM  OF  CASEIN 
PRODUCTION  OF  CASEIN  IN  THE  MILK  GLANDS 

Nissen  (1886)  observed  nuclear  changes  in  the  milk  gland  dur- 
ing secretion,  and  suggested  that  the  casein  may  form  by  union  of 
nuclein  from  the  nuclei  of  the  epithelial  cells  with  protein  from  the 
cytoplasm  of  the  gland  cells.  Michaelis  (1898)  added  support 
to  this  idea  by  finding  an  abundance  of  the  epithelial  cells  free  in 
the  lumen  of  the  alveoli  during  lactation;  and  Basch  (1898,  1903) 
isolated  a  nucleic  acid  from  the  gland,  from  which,  by  allowing  this 
substance  to  act  on  an  excess  of  blood  serum,  he  obtained  a  body 
having  so  many  of  the  properties  of  cow's  casein  that  he  felt  sure 
that  it  was  casein.  His  conclusion  was  that  within  the  alveoli  this  nu- 
cleic acid  from  the  cell  nuclei  (freed  either  by  destruction  of  the  cells 


PHOSPHORUS  METABOLISM  261 

or  by  an  active  .secretion  of  the  cells)  combines  with  transudated 
serum,  thus  forming  a  nucleoalbumin,  the  casein.  His  theory 
seemed  to  be  further  supported  in  that  he  was  unable  to  obtain 
purin  bases  from  his  nucleic  acid,  the  nucleic  acid  thus  correspond- 
ing with  the  paranucleic  character  of  casein;  but  the  nucleic  acid 
which  J.  A.  Mandel  and  Levene  (1905)  isolated  from  the  mammary 
gland  did  show  purin  bases.  This  tends  to  discredit  Basch's  theory, 
and  other  evidence  against  it  is  contributed  by  Lobisch's  (1906) 
studies  of  nuclein-protein  compounds  and  by  Borrino's  (1910,  1911) 
finding  a  nuclease  in  the  mammary  glands  present  only  during  lac- 
tation. This  nuclease  would  indicate  a  breaking  down  of  nuclein 
beyond  the  point  of  nucleic  acid ;  so  that  if  the  phosphorus  of  such  a 
compound  is  used  in  the  synthesis  of  casein  it  is  by  previous  destruc- 
tion of  the  acid. 

There  may  be  significance  in  the  coincidence  that  a  nucleopro- 
tein  which  Mandel  has  obtained  from  mammary  glands  yields  hydro- 
lytic  cleavage  products  in  approximately  the  same  proportions  as 
those  formed  from  casein. 

Certain  experiments  reported  in  1908  by  Michaelis  and  Rona 
(1908)  imply  a  relation  of  the  formation  of  casein  to  the  phenome- 
non of  milk-secretion,  the  significance  of  which  has  not  been  ex- 
plained. Subcutaneous  injection  of  casein  into  guinea  pigs  and  dogs 
was  followed,  in  females,  and  in  males  as  well,  by  a  swelling  of  the 
mammae,  and,  at  least  in  one  case,  by  the  production  of  true  milk. 
Examination  of  the  glands  showed  that  the  cells  had  undergone  the 
changes  usual  with  milk  secretion. 

Bergell  (1898b)  suggests  in  explanation  of  certain  figures  for 
the  phosphorus  of  the  blood  ash  of  lactating  female  animals,  in  com- 
parison with  the  normal  for  the  species,  that  a  higher  phosphorus 
content  of  the  blood  ash  may  be  associated  with  the  milk  producing 
function,  but  the  data  submitted  do  not  warrant  a  positive  assertion 
to  that  effect. 

DIGESTION  OF  CASEIN 

DIGESTION  IN  THE  STOMACH 

Rennet  action.  Casein,  unlike  other  proteins,  is  usually  intro- 
duced into  the  stomach  in  the  uncoagulated  state,  and  accordingly, 
a  special  coagulating  enzyme  is  provided  in  the  rennin.  Doubtless 
the  coagulation  is  a  favorable  preliminary  step  in  digestion.  The 
conditions  of  this  coagulation,  and  the  steps  of  the  process,  have 
been  discussed  in  the  consideration  of  the  chemistry  of  casein.  The 
casein  is  first  modified  to  paracasein,  and  then  precipitated  with 
calcium  salts.      The  same  action  may  be  brought  about  by  enzymes 


262  OHIO  EXPEEIMENT  STATION:  TECHNICAL  BUL.  5 

occurring  in  blood,  pancreas  and  other  organs.  That  of  the 
pancreatic  juice  is  probably  useful  with  milk  which  passes  from  the 
stomach  uncoagulated.  Whether  or  not  the  rennin  itself  has  any- 
further  action  on  casein,  under  favorable  circumstances,  is  uncer- 
tain (Van  Herwerden,  1907,  and  others).  Paracasein  is  found  to  be 
more  easily  and  more  thoroughly  digested  (in  vitro)  than  is  the  un- 
altered casein  (Hosl,  1910). 

Gaucher  has  reported  observations  on  the  time  taken  for  milk  and 
casein  to  pass  on  from  the  stomach,  and  the  physical  state  in  which 
the  casein  passes,  as  shown  by  duodenal  fistulas.  Gaucher  (1909) 
observed  three  stages  in  this  process  in  a  dog.  First,  the  milk 
passes  out  as  such  during  the  first  quarter  of  an  hour ;  second,  dur- 
ing the  second  quarter  of  an  hour  the  lactoserum  flows  out  mixed 
with  large  clots ;  and  third,  the  liquid  which  flows  later,  colored  by 
bile,  contains  fine  particles  of  coagulated  casein  held  in  suspension. 
Peptonizing  does  not  take  place  in  the  stomach.  Gaucher  (1911) 
made  similar  observations  on  a  child.  At  a  later  date  Gaucher 
(1912)  showed  with  a  dog  that  the  digestion  of  casein  in  the  con- 
tracted, solid  form  corresponds  only  to  the  third  of  these  stages. 
According  to  Gaucher  then — of  the  7  grams  of  casein  taken  in  250 
c.c.  of  milk,  4  grams  pass  the  pylorus  in  an  unaltered  form  during 
the  first  stage;  10-15  minutes'  secretion  of  gastric  juice  clots  the 
remainder  of  the  milk,  and  the  stomach  contractions  detach  rather 
coarse  fragments  of  the  clot,  1  gram  of  the  casein  passing  from  the 
stomach  in  this  stage ;  the  other  2  grams  contract  to  a  harder  mass, 
which,  as  the  muscular  action  of  the  stomach  becomes  more  and 
more  vigorous,  is  gradually  reduced  to  a  puree,  this  portion  leaving 
the  stomach  after  the  lapse  of  about  an  hour. 

Peptic  Digestion.  The  action  of  the  enzymes  of  the  digestive 
tract  has  been  studied  largely  by  artificial  digestion  in  vitro.  Study 
of  this  kind  with  artificial  gastric  juice  was  undertaken  in  1870  by 
Lubavin  (1870, 1877).  It  was  early  known  that  as  a  result  of  pep- 
tic digestion  there  pass  into  solution  albumoses  or  proteoses  (those 
of  casein  are  designated  caseoses)  and  peptones,  and  that  there  re- 
mains an  insoluble  nuclein-like  body  richer  in  phosphorus  than  the 
casein.  This  body  has  been  called  pseudonuclein  or  paranuclein, 
and  it  has  been  shown  that  under  favorable  circumstances  it  may  be 
itself  digested  by  the  pepsin-hydrochloric  acid  mixture.  On  fur- 
ther digestion  it  is  said  to  become  more  rich  in  phosphorus  (von 
Szontagh,  1892,  1893).  References  to  investigations  in  this  field 
were  given  in  connection  with  the  consideration  of  comparisons  of 
different  caseins  in  this  respect. 


PHOSPHORUS  METABOLISM 


263 


According  to  Chittenden  and  his  associates,  (Chittenden  and 
Painter,  1885 ;  Jackson,  1900)  the  caseoses  contain  no  organic  phos- 
phorus, but  the  paranuclein  (which  they  call  dyspepton)  always  con- 
tains over  2  percent  of  phosphorus  in  organic  combination.  A  small 
part  of  the  phosphorus  goes  into  solution  as  inorganic  phosphoric 
acid. 

Clara  Willdenow  (1893)  conducted  peptic  digestion  experiments 
on  casein  (in  vitro),  and  found  split  off  a  phosphorus-containing 
body,  which  contains  its  phosphorus  mainly,  not  as  calcium  phos- 
phate, but  in  organic  combination,  agreeing  with  Lubavin,  but  not 
with  Chittenden.  This  substance,  from  its  properties  and  phos- 
phorus content,  seemed  like  nuclein  or  nucleic  acid.  Willdenow  ob- 
tained but  a  single  body  containing  phosphorus  and  sulphur,  not 
like  Lubavin's  finding  of  two  such  separable  by  sodium  carbonate. 

Von  Moraczewski  (1895a),  under  Drechsel's  direction,  studied 
the  distribution  of  the  phosphorus  between  nuclein  and  the  filtrate 
from  the  same  after  the  digestion  of  casein  with  pepsin.  Various 
changes  of  the  conditions  of  the  digestion  were  made,  especially  as 
to  the  concentration  of  the  solution  and  the  duration  of  digestion. 
The  results  given  below  are  taken  from  the  author's  table : 

PHOSPHORUS  OF  THE  NUCLEIN  RESULTING  FROM  PEPTIC 
DIGESTION  OF  CASEIN 


Concentra- 

Weight of  nu- 

Nuclein P 

tion  of  case- 

Time of 

vVeigtit  of 

Weight  of 

clein  from  100 

to  100  gm. 

Phosphorus 

Exp. 

in  solution 

digestion 

pepsin 

casein 

gm.  casein 

casein  P 

in  nuclein 

No. 

Percent 

Days 

Grams 

Grams 

Grams 

Grama 

Percent 

I 

4 

8 

0.44 

14.26 

13.60 

34.20 

2.10 

n 

4 

5 

1.38 

8.15 

14.72 

37.50 

2.40 

III 

4 

3 

1.39 

10.19 

17.40 

44.20 

2.19 

I 

4 

2 

0.5 

3.5 

14.84 

50.54 

0.88 

I 

6 

14 

0.5 

3.00 

3.60 

20.66 

3.87 

II 

6 

10 

0.5 

3.00 

6.67 

29.20 

3.13 

III 

6 

7 

0.5 

3.00 

8.31 

31.31 

2.70 

I 

0.7 

5 

1.0 

5.27 

1.29 

6.75 

4.10 

II 

0.7 

1 

1.0 

2.68 

4.64 

18.20 

3.15 

III 

1.44 

7 

1.0 

5.46 

5.96 

42.61 

6.86 

IV 

1.44 

5 

1.0 

9.11 

8.37 

26.17 

3.43 

I 

3.45 

2 

1.5 

33.80 

16.48 

44.51 

2.08 

II 

3.45 

10 

1.5 

33.80 

18.17 

52.59 

2.27 

III 

3.51 

10 

2.0 

31.61 

17.99 

56.40 

2.44 

IV 

3.51 

2 

1.0 

31.61 

21.10 

63.21 

2.82 

It  was  concluded  that :  "The  casein  does  not  have  all  of  its  phos- 
phorus in  the  form  of  nuclein,  for  the  amount  found  ranges  from  6 
to  60  percent,  but  is  never  the  total  phosphorus."  "Nuclein  from 
cow's  milk  does  not,  even  after  long  digestion,  go  completely  into  so- 
lution      The  amount  is  affected  by  the  time  of  digestion 


264 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


and  still  more  by  dilution.  In  very  dilute  solution  the  amount  of 
nuclein  is  very  small  and  it  is  very  rich  in  phosphorus.  With  longer 
time  of  digestion  under  these  circumstances  the  amount  of  nuclein 
decreases  and  its  phosphorus  content  increases.  In  concentrated 
solutions  nuclein  precipitates  in  large  amount  and  loses 
little  phosphorus  even  on  protracted  digestion."  "In  the  di- 
gestion fluid  the  phosphorus  can  be  precipitated  by  magnesia  mix- 
ture directly  only  after  long  digestion  and  in  very  dilute  solution. 
The  degree  of  dilution  plays  a  greater  part  than  the  amount  of  pep- 
sin or  the  time  of  digestion  here  also "  Von  Moraczewski's 

theory  is  that  the  nuclein  or  nucleic  acid  from  it  carries  down  with 
it  some  protein  body  such  as  unaltered  casein  or  caseoses. 

Sebelien  (1895)  found  44  and  33  percent  of  the  phosphorus  of 
casein  in  the  paranuclein  portion. 

Salkowski  reported  experiments  (Salkowski,  1893a,  1893b ;  Sal^ 
kowski  and  Hahn,  1894-95)  from  which  he  felt  sure  that  in  peptic 
digestion  the  main  part  of  the  phosphorus  of  casein  remains  in  solu- 
tion in  some  kind  of  organic  combination,  only  a  smaller  part  being 
in  the  insoluble  paranuclein.  It  was  further  found  that  the  amount 
of  phosphorus  in  the  paranuclein  varied  with  the  conditions  of  di- 
gestion, the  more  unfavorable  the  conditions,  the  greater  the  quan- 
tity of  paranuclein  and  the  greater  also  the  quantity  of  phosphorus 
in  the  insoluble  product.  The  following  table  is  taken  from  the  ar- 
ticle by  Salkowski  and  Hahn. 

RESULTS  OF  PEPSIN  DIGESTION  OF  CASEIN— Percent 


Exp. 

Paranu- 
clein 
portion 

Albumose 
portion 

Phosphorus  con- 
tent of 

Percent  of  total 
phosphorus  in 

Paranu- 
clein 

Albu- 
mose 

Paranu- 
clein 

Albu- 
mose 

II 
III 
IV 

V 
VI 

6.8 
6.76 
18.5 
15.2 
21.05 

93.2 

93.24 

81.5 

84.8 
78.95 

2.41 

0.55  (?) 
2.27 
2.18 
2.11 

0.74 
0.87  (?) 
0.59 
0.58 
0.51 

19.0 
4.3  (?) 

41.2 
41.9 
52.5 

81.0 
95.7  (?) 

58.5 
58.1 
47.5 

r  Conditions  most  favorable. 

(.Too  little  of  pepsin  solution, 
f        or  pepsin  less  active. 

Salkowski  (1896a)  reported  the  conditions  most  favorable  to 
complete  digestion,  and  later  (1899)  stated  that  by  carrying  on  the 
digestion  under  retarding  circumstances  he  was  able  to  identify  an 
intermediary  phosphorus-containing  albumose.  He  says:  "The  pep- 
sin digestion  of  casein,  then,  proceeds  in  three  stages:  1.  The 
transformation  of  the  casein  into  an  albumose,  2.  splitting  off 
of  paranuclein  from  this,  3.  complete  solution  of  the  paranuclein 
and  further  digestion  of  the  albumose.  While  it  is  not  possible 
sharply  to  separate  the  second  and  third  stages  from  one  another, — 
as  during  the  splitting  off  of  the  paranuclein a  part  at  least 


PHOSPHORUS  METABOLISM  265 

of  it  goes  into  solution, — the  distinction  between  the  first  and  sec- 
ond stages  is  easy,  as  the  splitting  off  of  the  paranuclein  takes  a  def- 
inite time,  and  moreover  the  process  can  be  interrupted  before  all 
the  casein  has  changed  to  albumose." 

Zaitchek  (1904)  and  von  Szontagh  (1905),  under  Tangl's  direc- 
tion, made  comparative  digestion  experiments  on  casein  from  dif- 
ferent kinds  of  milk,  the  results  of  which  are  evident  in  the  conclu- 
sions which  we  quote  from  Zaitchek. 

"1.  By  pepsin-hydrochloric  acid  digestion  experiments  we  de- 
termined that  woman's,  ass's  and  mare's  milk  is  completely  digested, 
while  the  casein  of  cow's,  goat's  and  buffalo's  milk  under  the  same 
conditions  of  experiment  (temperature  38°C,  time  72  hours)  is  only 
8, 14  and  15  percent  soluble. 

"2.  Each  kind  of  milk  which  is  not  soluble  in  pepsin-hydro- 
chloric acid  without  residue  leaves  a  different  amount  of  pseudonu- 
clein  from  the  casein  obtained  from  it.  The- latter  gave  without  ex- 
ception 2-3  percent  smaller  pseudonuclein  residue  than  the  milk 
containing  the  same  amount  of  casein.  The  casein  precipitated 
from  woman's,  ass's  and  mare's  milk  is  as  completely  soluble  as  the 
milk  itself. 

"3.  The  woman's,  ass's  and  mare's  milk  not  only  contains  an 
absolutely  smaller  amount  of  casein  than  cow's,  goat's  and  buffalo's 
milk,  but  a  smaller  portion  of  the  total  nitrogen,  also,  is  in  the 
casein. 

"4.  Under  like  conditions  of  experiment  the  different  kinds  of 
pure  casein  give  different  amounts  of  pseudonuclein  (0-15  percent) . 

"5.  The  addition  of  thymol,  toluol  and  chloroform  hinders  the 
casein-dissolving  action.  The  checking  effect  increases  with  the 
amount  added. 

"6.  Both  the  concentration  relations  and  the  duration  of  ac- 
tion of  the  pepsin-hydrochloric  acid  have  considerable  influence  oh 
the  solubility  of  casein  in  pepsin-hydrochloric  acid. 

"7.  The  drying  of  casein  at  110°C.  considerably  reduces  the 
solubility  in  pepsin-hydrochloric  acid." 

Some  of  Robertson's  studies  (1906-07, 1907, 1909a,  1910b,  1911, 
Robertson  and  Biddle,  1911)  on  the  reversible  action  of  enzymes 
having  been  made  with  casein,  this  work  and  his  theory  regarding 
the  matter  may  be  considered  in  this  connection. 

One  of  the  stages  in  peptic  digestion  of  casein  is  quite  marked 
because  of  the  difficultly  soluble  nuclein  (or,  better,  "pseudo-"  or 
"paranuclein")  produced.  It  has  been  thought  that  digestion  of  the 


266  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

casein  from  cow's  milk  could  not  be  carried  beyond  this  point  by  pep- 
sin, but  it  has  since  been  shown  that  under  favorable  conditions  the 
paranuclein  may  be  made  entirely  soluble.  Now,  Robertson  has 
succeeded,  by  the  use  of  pepsin,  in  bringing  about  a  synthesis  of 
paranuclein  (or  one  of  the  constituents  of  the  mixture  usually  ob- 
tained as  paranuclein)  from  the  products  of  complete  peptic  diges- 
tion of  casein.  The  obvious  explanation  is  that  pepsin  acts  as  a  cata- 
lytic agent,  accelerating  the  opposite  phases  of  a  reversible  reaction, 
the  breaking  down  of  casein  into  the  simpler  products,  and  the  union 
of  the  simpler  products  toward  the  formation  of  casein.  Robertson's 
theory  modifies  this  interpretation  a  little  in  that  he  thinks  that 
there  are  two  forms  of  the  enzyme  active  in  the  two  processes.  Just 
as  the  cleavage  products  differ  from  the  protein  by  the  elements  of 
water,  and  may  be  looked  upon  as  successively  more  and  more  hy- 
drated,  so  the  enzyme  is  supposed  to  have  two  forms,  the  hydrated 
and  the  anhydrous.  The  hydrated  form  is  effective  in  introducing 
water  into  the  protein  (accelerating  hydrolysis),  and  the  anhydrous 
form,  in  removing  water,  so  that  the  simpler  bodies  unite  with  one 
another  (accelerating  synthesis).  The  theory  is  that  the  effect 
is  brought  about  by  the  aid  of  a  temporary  union  of  the  ferment 
with  the  protein.  Representing  the  two  forms  of  the  ferment  by 
HFFOH  and  FF,  and  the  protein  and  its  cleavage  products  by 
HXXOH  and  HXOH,  the  types  of  reaction  concerned  in  the  process- 
es under  discussion  may  be  shown  as  follows : 

HXXOH+HFFOH^±HXXFFOH+H20  (1) 

HXXFFOH+H2CM2HXOH+FF  (2) 

FF+H2O^HFFOH  (3) 

In  case  of  hydrolysis  (1)  represents  the  union  of  protein  with 
the  hydrated  ferment;  (2),  the  breaking  up  of  this  union,  leaving 
the  protein  split,  and  the  ferment  in  its  anhydrous  form;  (3),  the 
ferment  again  taking  up  water  to  itself  from  the  medium.  In  syn- 
thesis the  processes  are  reversed  under  the  influence  of  the  dehy- 
drated enzyme.  The  ratio  of  the  velocity  of  the  reaction  in  the  two 
directions  is  affected  by  the  concentration  of  the  protein  or  its 
cleavage  products  and  of  the  enzyme,  by  the  temperature,  and  prob- 
ably also  by  the  alkalinity  or  acidity ;  and  this  ratio  determines  the 
final  equilibrium. 

Bayliss  (1913)  believes  that  Robertson's  (1907)  synthesis  of 
paranuclein  from  casein  digestion  products  by  the  action  of  pepsin  is 
not  a  synthesis  at  all,  but  a  colloidal  precipitation  without  connec- 
tion with  the  enzyme. 


PHOSPHORUS  METABOLISM  267 

Kuttner  (1909)  reports  a  study  of  peptic  digestion  of  casein 
from  the  standpoint  of  the  acidity  of  its  cleavage  products.  Diges- 
tions were  made  with  differences  in  proportion  of  gastric  juice 
(from  dog) ,  in  acidity,  in  dilution,  in  temperature  and  in  duration  of 
digestion,  with  phosphorus  and  nitrogen  determinations  on  the  dis- 
solved and  undissolved  portions,  and  in  one  series  on  the  alcohol  pre- 
cipitable  portion  of  the  products  of  digestion.  During  one  series 
the  digestive  products  were  removed  at  stated  intervals,  and  fresh 
mixture  substituted.  In  each  case  the  digestion  was  stopped  at  the 
time  that  a  precipitation  of  the  products  of  digestion  appeared.  The 
nature  of  the  data  is  evident  from  the  table  on  p.  268.  It  will  be  ob- 
served that  a  considerable  increase  of  acidity  occurs  during  diges- 
tion. Of  this  a  certain  fraction  is  spoken  of  as  the  non-salt-precip- 
itable  acidity,  meaning  the  fraction  of  the  final  acidity  which  is  due 
to  some  compound  not  precipitated  by  thirty  percent  sodium  chlor- 
ide solution. 

The  author's  conclusions  are :  "There  is  every  reason  to  believe 
that  casein  is  cleaved  during  peptic  digestion  into  a  phosphorus- 
free  and  a  phosphorus-containing  portion.  While  cleavage  of  the 
former  continues  even  under  the  most  unfavorable  digestive  con- 
ditions, the  latter  (the  paranuclein)  is  less  easily  digestible,  though 
its  further  digestibility  is  such  that  even  under  the  most  unfavor- 
able conditions  a  fraction  is  always  attacked.  By  the  further  diges- 
tion of  paranuclein  under  favorable  conditions  there  results 
a  peptically  indigestible  fraction  (Kiihne's  anticomplex) , 
while  the  phosphorus-containing  constituent  which  is  separated 
from  it  by  peptid  cleavage  is  further  cleaved  into  more  and  more 
highly  phosphorized  acid-like  compounds  (paranucleic  acids) .  These 
are  in  the  main  responsible  for  the  increase  in  acidity  of  the  digest- 
ive products  observed  during  the  digestion  of  casein." 

According  to  the  observations  of  Miss  Goldthwaite  (1910),  the 
presence  of  carbohydrates  retards  the  peptic  digestion  of  casein,  and 
by  an  amount  proportional  to  the  amount  of  carbohydrate  present. 

Long  (1907a)  made  protracted  digestions  (nearly  two  months) 
of  casein  from  cow's  and  from  goat's  milk  observing  the  "free"  and 
"total"  acid  in  the  digestion  mixtures  as  digestion  progressed,  the 
electrical  conductivity  of  this  solution,  the  weights  of  undissolved 
and  dissolved  solids,  and  the  conductivity  and  acidity  of  this  dis- 
solved solid  on  being  redissolved  after  evaporation.  During  diges- 
tion the  electrical  conductivity  values,  and  the  total  and  free  acidity 
varied  regularly.  Both  acidity  values  were  lower  for  the  goat 
casein  than  for  cow's  casein.  A  larger  residue  of  pseudonuclein  was 
finally  left  from  the  casein  of  goat's  milk ;  the  dissolved  substances 
were  thought  to  be  similar  in  nature. 


268 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


ft 

O 

co 
o 


O 

o 

ft" 
ft 
ft 


ft 

CO 

3 

ft 
O 

o 

H 
CO 

ft 


H 

Oh 
ft 


■a  s. ■§ 


«4-(    LQ 


o  QJ 

5  a 


a  n 


rt^  in 
g  ™N, 


OINt»000-*CO-« 


in  cio 

CDCDOOOOOCOCiaO 

"71  tl  "71 TJ  Tl  t!  T!  "tJ 

oooooooo 


OOOOOOOO 


CO?OOf-"*CDCOCO 
*— I001OO3001OCOO3 

cocM'iric^ooooTn'co 

CM-tflLQCOCPC-OOOO 


rocooomcoocoo) 
io»-Hoicocococcio 

I-ICMCNCOCQ-^IIOIO 


TH— l-*0000(M1030 

COCOCOi-HCC'JtOTOO 

»— ir~mODi>iascNico 
N  CO  lOlQCfi  CCISCo 


masCOCNKNOOTHOJ 
COCDCOOOCDlOOrH 

odcNiTtior^ot^co 

C010eoCOCMCNi-1i-< 


•OTCOi— IOCOOS 

•eoco^'miQio 


COCOIOCDOSO^CO 

cdos'^'^ojo^t-J 

i-l(MW>SllOt^CC05 


COJN 

IQrH 


5288SS 


oooiooioinin 

rH-Hf-IOJOJtMCN 


PHOSPHORUS  METABOLISM 


269 


DIGESTION  IN  THE  INTESTINE 

In  the  intestine  the  casein,  whether  still  unaltered,  or  changed 
by  the  pepsin  to  proteoses  and  peptones,  can  be  further  split  by  the 
trypsin  of  the  pancreatic  juice  and  the  erepsin  of  the  intestinal  se- 
cretion (Hammarsten,  1911).  The  paranuclein  from  casein  is  also 
actively  digested  by  trypsin. 

Brugsch  and  Masuda  (1911)  state  that  the  intestinal  juice  can 
not  digest  casein,  though  an  extract  of  the  colon  bacillus  does  so,  and 
that  the  feces  do  not  possess  a  casein-cleaving  capacity  if  the  pan- 
creatic secretion  be  excluded. 

The  absorption  of  the  cleavage  products  of  casein  takes  place 
at  least  principally  as  amino  acids.  The  form  in  which  the  phos- 
phorus of  casein  is  absorbed  has  not  been  determined. 

Artificial  Digestion  Tests.  Sir  William  Roberts  (1879,  1881) 
first  noticed  the  curdling  of  milk  by  pancreatic  extract ;  also  that  the 
first  principal  phase  of  the  tryptic  digestion  of  casein  is  the  forma- 
tion of  a  modification  of  the  casein,  which  he  and  others  since  have 
called  metacasein.  In  both  of  these  respects  the  action  is  very  sim- 
ilar to  that  of  rennet,  as  others  have  noted  (Edkins,  1891 ;  Halli- 
burton and  Brodie,  1896).  Harris  and  Gow  (1892)  found  the  ac- 
tion lacking  in  the  pancreas  of  some  animals. 

Von  Szontagh  (1894)  states  that  on  digestion  with  pancreatic 
juice  the  casein  is  first  entirely  dissolved,  and  then  a  precipitate 
forms.  In  an  alkaline  solution  the  precipitate  is  less,  but  evidently 
because  of  a  solvent  action  of  the  alkali,  and  not  because  of  digestion 
of  the  nuclein. 

Sebelien  (1895)  reports  digestion  experiments  with  pan- 
creatin-soda  solution.  Below  are  the  author's  figures  for  his  fourth 
and  fifth  series  of  experiments,  in  which,  with  two  different  concen- 
trations of  casein  solution,  increasing  amounts  of  pancreatin  solu- 
tion were  used  (the  total  volume  of  digesting  solution  being  kept  at 
300  cc.  by  regulation  of  the  volume  and  concentration  of  the  soda 
solution  added) .     In  this  case  the  residue  precipitated  on  acidifying 

DEGREE  OF  DIGESTION  OF  CASEIN  AND  ITS  PHOSPHORUS  BY 

PANCREATIN 


Time  of  digestion 

Casein  solution 

Pancreatin               Soda 
solution               ._ 

Percent 

Casein 
undissolved 

Percent 

Phosphorus 
undissolved 

Percent 

18  hours 

200  cc,  2.076* 
200  cc 
200  cc 

10  cc,     5* 
20  cc 
40  cc 

0.167 
0.167 
0.167 

1.2 

2.3 
2.5 

2.2 

2.3 
2.3 

200 cc,  .556* 
200  cc 
200  cc 

25  cc,     1* 
60  cc 
75  cc 

0.208 
0.208 
0.208 

2.5 
3.5 
2.5 

2.1 

M        «  • 

1.6 
1.2 

270  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

the  solution  after  a  certain  time  of  digestion  was  weighed,  and 
tested  for  phosphorus.  This  residue  was  considered  to  be  undis- 
solved casein.  It  was  but  a  small  percent  of  the  casein  by  weight, 
and  was  not  like  the  pseudonuclein,  which  forms  a  much  larger  part 
of  the  casein.  As  the  figures  show,  the  casein  and  the  casein  phos- 
phorus were  practically  completely  digested. 

Rotondi  (1902)  reports  that  casein,  from  either  human  or 
bovine  milk,  is  much  more  readily  digested  than  fibrin,  by  both  pep- 
sin-hydrochbric  acid  and  pancreatin,  the  digestion  by  pancreatin 
being  especially  easy. 

E.  Fischer  and  Abderhalden  (1903)  say  that  more  hydrolysis  of 
casein  takes  place  by  the  combined  working  of  pepsin-hydrochloric 
acid  and  pancreatin  than  by  pancreatin  alone. 

Biffi  (1898), using  a  pancreatic  ferment  prepared  from  beef  pan- 
creas, concluded  that  under  favorable  conditions  casein  is  completely 
digested  by  that  enzyme,  about  4  percent  splitting  off  as  tyrosin, 
with  the  formation  of  primary  and  secondary  albumoses  and  an  anti- 
peptone  which  correspond  to  those  of  fibrin.  All  of  these  products 
are  probably  free  from  phosphorus ;  but  in  the  soluble  product  phos- 
phorus is  always  present  in  two  forms,  a  part  being  directly  precip- 
itable  by  magnesia  mixture  and  another  part  being  so  precipitable 
only  after  fusion  with  soda  and  potassium  nitrate.  The  phosphoric 
acid  increases  with  the  duration  of  digestion  and  with  the  amount  of 
ferment,  at  the  expense  of  the  organic  phosphorus.  The  organic 
can  further  be  changed  into  inorganic  by  the  action  of  dilute  alkali 
solution  or  by  boiling  with  barium  carbonate. 

From  the  tube  digestions  made  by  Rachford  (1900)  it  may  be 
said  that  pancreatic  digestion  of  the  casein  of  cow's  milk  is  facili- 
tated by  the  presence  of  maltose  solution,  of  lime  water,  or  of  0.4 
percent  sodium  carbonate  solution.  Hydrochloric  acid  was  found 
very  slightly  to  retard  the  action  of  pancreatic  juice  alone,  but 
greatly  to  increase  the  action  in  the  presence  of  bile.  "Bile  assists 
the  pancreatic  juice  in  the  digestion  of  casein,  but  it  renders  even 
greater  assistance  when  the  casein  is  partly  saturated  with  hydro- 
chloric acid." 

Plimmer  and  Bayliss  (1906)  studied  the  action  of  enzymes  and 
of  alkali  on  casein.      They  summarize  their  results  as  below: 

"1.  The  whole  of  the  phosphorus  of  caseinogen,  except  for  a 
very  small  residue,  is  converted  into  a  soluble  form  by  the  action  of 
trypsin  in  24  hours. 

"2.  The  curve  of  its  rate  of  separation  is  exactly  parallel  to  the 
curve  of  the  electrical  conductivity  (see  Bayliss,  1904)  during  the 
first  7-8  hours ;  its  rate  of  separation  after  this  time  is  less  rapid. 


PHOSPHORUS  METABOLISM  271 

"3.  The  small  insoluble  residue  is  partly  derived  from  the 
trypsin,  and  partly  from  the  caseinogen,  and  consists  most  probably 
of  the  products  of  decomposition  of  nucleoproteid. 

"4.  The  'soluble  P205'  consists  of  inorganic  phosphoric  acid — 
35  percent —  and  organic  phosphorus,  65  percent. 

"5.  The  phosphorus  of  caseinogen  is  very  slowly  converted  in- 
to a  soluble  form  by  the  action  of  pepsin.  The  quantity  thus 
changed  is  only  70  percent  in  149  days,  and  consists,  except  for  a 
negligible  quantity  of  inorganic  phosphoric  acid,  of  organic  phos- 
phorus. 

"6.  Papain,  in  its  action  on  caseinogen,  is  intermediate  in 
power  between  pepsin  and  trypsin.  In  its  rate  of  splitting  off  of 
'soluble  P2(V  and  'soluble  nitrogen'  it  resembles  trypsin  the  more 
closely,  but  it  is  much  slower. 

"It  acts  best  in  a  neutral  or  faintly  acid  medium ;  its  action  is 
slower  in  a  slightly  alkaline  medium  and  is  almost  inhibited  by  0.5 
percent  sulphuric  acid  and  0.5  percent  sodium  carbonate. 

"7.  Ovovitellin,  containing  lecithin,  is  very  slowly  digested  by 
trypsin  in  comparison  with  caseinogen.  Only  one-half  of  its  phos- 
phorus is  converted  into  a  soluble  form  in  36  days.  A  similar 
quantity  is  probably  contained  in  the  lecithin  portion  of  the  mole- 
cule. 

"8.  One  percent  caustic  soda  converts  the  whole  of  the  phos- 
phorus of  caseinogen  into  'soluble  P205'  in  24  hours.  It  resembles 
trypsin  very  closely  in  its  rate  of  action. 

"9.  The  'soluble  P205'  produced  by  one  percent  caustic  soda 
in  24  hours  consists  entirely  of  inorganic  phosphoric  acid.  In  the 
same  time  the  quantity  of  'soluble  nitrogen'  scarcely  increases. 

"10.  The  organic  phosphorus  produced  by  the  action  of  tryp- 
sin on  caseinogen  is  not  completely  converted  into  inorganic  phos- 
phoric acid  by  the  action  of  one  percent  caustic  soda.  The  total 
quantity  of  inorganic  phosphoric  acid  produced  by  trypsin  and  sub- 
sequently by  one  percent  caustic  soda  is  50  percent  of  the  total  phos- 
phorus of  caseinogen." 

Robertson  (1906-7),  from  his  study  x>f  tryptic  digestion  of  case- 
in in  relation  to  the  ion-proteid  theory,  concludes  (in  part)  that  neu- 
tral casemates  in  solution  undergo  fairly  rapid  autohydrolysis ;  that 
the  velocity  of  hydrolysis  of  calcium  caseinate  by  trypsin  at  mod- 
erate substrate  concentration  is  directly  proportional  to  the  amount 
of  trypsin  present — at  higher  substrate  concentrations  the  rate  of 
increase  being  a  little  greater.  A  variety  of  salts  (including  di- 
sodium  phosphate)  were  found  to  accelerate  in  different  degrees  the 


272  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

velocity  of  the  hydrolysis  of  calcium  and  sodium  caseinates ;  mono- 
sodium  phosphate  had  a  retarding  influence.  The  special  study  of 
the  part  played  by  the  alkali  in  the  hydrolysis  of  proteins  by  tryp- 
sin (Robertson  and  Schmidt,  1908),  the  alkalinity  being  followed  by 
means  of  the  gas-chain,  brings  out  facts  against  the  view  that  the 
OH"  ions  in  a  tryptic  digest  play  the  part  of  an  accessory  catalyzer, 
though  it  is  suggested  that  the  real  catalyzer  in  these  systems  may 
be  a  hydrolyzable  compound  of  trypsin  with  the  base  present.  The 
investigations  of  Walters  (1912a,  1912b)  are  a  continuation  of  that 
of  Robertson. 

ANIMAL  EXPERIMENTS  WITH  CASEIN 

Sandmeyer  (1895),  testing  the  possibility  of  the  absorption  of 
the  paranuclein  from  casein,  introduced  such  paranuclein  into  a  dog 
by  means  of  a  probe,  and  judged  of  its  absorption  by  phosphorus  de- 
terminations in  the  urine.  If  one  may  draw  any  inference  from  such 
an  observation,  something  more  than  ^  of  the  paranuclein  was  ab- 
sorbed. The  dog  received  only  water  for  two  x)Y  three  days  before 
the  introduction  of  the  phosphorus  compound,  and  the  urine  showed 
0.33-0.37  grams  P205  per  day;  on  the  paranuclein  days  2.864  and 
4.296  grams,  respectively,  of  P205  were  introduced,  and  caused  1.34 
and  1.84  grams  to  appear  in  the  urine ;  in  each  case  on  the  day  fol- 
lowing the  experiment  the  urinary  phosphorus  returned  to  its  pre- 
vious value  (0.33  and  0.30  grams  P205). 

Salkowski  (1901)  fed  the  iron  compound  of  his  so-called  para- 
nucleic  acid  to  rabbits.  Absorption  occurred,  as  judged  on  the  evi- 
dence of  finding  the  iron  content  of  the  livers  of  the  rabbits  which 
received  this  iron  compound  appreciably  above  that  of  those 
which  had  a  corresponding  diet  without  this  substance,  and  above 
that  of  those  which  received  iron  in  the  form  of  its  combination 
with  atmidalbumose  (an  albumose  derived  from  fibrin),  or  as  fer- 
ratin. 

Another  study  on  the  absorption  of  products  of  the  decomposi- 
tion of  casein  is  that  of  E.  Voit  and  Zisterer  (1909-10)  on  dogs. 
They  compared  the  nitrogen-sparing  value  of  undigested  casein  with 
that  of  the  products  of  pancreatin  digestion,  and  those  of  acid  hy- 
drolysis of  casein.  ,The  conclusion  is  that  the  physiological  value  of 
protein  bodies  is  unfavorably  influenced  by  a  deep  cleavage,  and 
that,  in  general,  protein  bodies  do  not  undergo  complete  cleavage  in 
the  digestive  tract,  but  that  certain  complexes  are  absorbed  un- 
changed. No  observations  were  made  with  regard  to  the  phospho- 
rus of  the  casein  or  its  decomposition  products. 

These  conclusions  are  almost  identical  with  those  of  Abder- 
halden  and  Rona  (1904,  1905)  from  their  observations  as  to  main- 
taining life  in  mice  and  meeting  the  nitrogen  requirement  of  a  dog. 


PHOSPHORUS  METABOLISM 


273 


With  regard  to  the  mice,  the  report  is  that  the  products  of  pancre- 
atin  digestion  of  casein  serve  about  as  well  as  pure  casein,  while 
those  from  the  combined  digestion  by  pepsin  and  pancreatin  (fur- 
ther split)  are  of  less  value.  Mice  fed  with  the  latter,  however, 
live  longer  than  starving  mice,  while  those  fed  with  the  products 
of  acid  hydrolysis  do  not.  The  results  with  the  dog  were  of  like 
import.  In  later  work,  however,  Abderhalden  (1912)  showed  that 
dogs  are  able  to  maintain  nitrogen  equilibrium  and  retention  on 
products  of  either  acid  or  enzymatic  splitting  of  proteins,  or  on  a 
mixture  of  recognized  amino-acids. 

Marcuse  (1896, 1897)  carried  out  nitrogen  and  phosphorus  bal- 
ance experiments  with  dogs  on  a  mixed  diet  in  which,  in  some  cases, 
the  protein  given  was  in  the  form  of  casein,  and  in  some  cases  in 
the  form  of  flesh.  In  either  case  a  salt  mixture  including  phos- 
phates was  given  with  the  other  food,  and  the  conditions  were  gen- 
erally too  complex  for  definite  inferences ;  but,  on  the  whole,  both 
nitrogen  and  phosphorus  retention  were  greater  for  the  casein  peri- 
ods than  for  the  flesh  periods. 

Schreiber  and  Waldvogel  (1897)  report  15  cases  of  various 
diseases  in  which  "Sanose,"  a  protein  preparation  containing  80  per- 
cent casein  and  20  percent  albumose,  was  taken  with  the  food  or 
used  in  its  preparation.  It  was  found  useful  in  cases  where  meat 
was  either  a  distasteful  or  an  undesirable  article  of  diet.  Urine  an- 
alyses are  submitted. 

Knoepfelmacher  (1898a,  1898b,  1899,  1900)  studied  the  use  of 
the  casein  of  milk  by  infants  and  older  children  by  a  comparison  of 
the  nitrogen  and  organic  phosphorus  of  their  feces  with  the  casein 
nitrogen  and  phosphorus  ingested.  The  methods  used  in  the  earli- 
est work,  comparing  the  use  of  mother's  milk  and  cow's  milk,  he 
says  later  were  faulty.  The  following  table  from  Knoepf elmacher's 
latest  work  shows  no  important  percentage  differences  in  the  diges- 
tion of  either  the  nitrogen  or  phosphorus  of  casein  by  infants  and 
older  convalescent  children. 

NITROGEN  AND  PHOSPHORUS  OUTGO  IN  THE  FECES  FROM  A  DIET 
OF  COW'S  MILK— INFANTS  AND  OLDER  CHILDREN 


Age  of  child 


Length 
of  ex- 
peri- 
ment 

Days 


Intake 


Milk 


Casein 
Grams 


Casein  N 
Grams 


Casein  P 
Grams 


Outgo  in  feces 


N 
Grams 


Total  P 
Grams 


Organic  P 
Grams 


11  yrs . . . 
8  yrs . . . 

7  yrs... 
0i  mos 

5  mos. . 

6  mos. . 

8  mos. . 


13,250 
10,681 
8,815 
4,770 
3,498 
4.400 


296.19 
341.01 
277.64 
112.21 
79.56 
60.52 
73.91 


46.498 
53.538 
43.5913 
17.6169 
12.49 
9.683 
11.825 


2.517 

2.898 

2.3599 

0.9537 

0.6762 

0.514 

0.628 


3.6088 

3.54 

1.723 

1.238 

0.928 

0.664 

0.6513 


8.33 

6.68 

3.645 

1.149 

0.914 

0.478 

0.856 


0.1225 

0.12 

0.1229 

0.0526 

0.0237 

0.0264 

0.0343 


274  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Prausnitz  and  his  associates  (Micko,  Miiller,  Poda  and  Praus- 
nitz,  1900;  Poda  and  Prausnitz,  1900;  Micko,  1900;  Miiller,  Paul, 
1900)  made  similar  feces  studies  with  older  people.  In  the  intro- 
ductory article  the  whole  work  is  summarized,  with  a  discussion  of 
the  general  conclusion  that  the  feces  are  made  up  in  the  main  of 
residues  from  the  digestive  juices,  rather  than  of  food  residues,  the 
amount  of  such  juices  being  modified  by  the  kind  of  food  taken. 
Poda  and  Prausnitz  concluded  that  casein,  in  the  form  of  the  prep- 
aration called  "Plasmon,"  was  utilized  as  well  by  healthy  men  as  the 
flesh  with  which  it  was  compared.  Micko  looked  for  paranuclein 
and  for  nuclein  in  the  feces.  Both  were  found  in  the  feces  from 
both  these  kinds  of  diet,  and  from  ordinary  mixed  diet ;  of  each  the 
plasmon  feces  showed  less  than  the  flesh  feces.  Paul  Miiller  con- 
cluded that  a  diet  of  cow's  milk  leaves,  with  neither  infants  nor 
adults,  a  phosphorus-rich  casein  residue  in  the  feces. 

Cronheim  and  Erich  Miiller  (1900,  1902)  compared  the  useful- 
ness, to  children,  of  lecithin  and  casein,  by  feeding  milk-powder  with 
and  without  added  egg-yolk.  The  periods  were  very  short ;  the  re- 
sults were  variable,  and  the  observations  are  not  very  significant. 
In  these,  and  in  similar  feeding  experiments  on  dogs  and  on  guinea 
pigs,  there  was  thought  to  be  somewhat  better  storage  of  nitrogen 
from  lecithin-containing  food  than  from  casein. 

Comparisons  of  Raw  and  Sterilized  Milk.  Jemma  (1899)  re- 
ports from  his  comparison  of  the  action  of  enzymes  on  sterilized 
and  unsterilized  milk  that  pepsin-hydrochloric  acid  digests 
unsterilized  milk  more  quickly  than  sterilized,  while  both 
pancreatin  and  rennin  digest  the  sterilized  more  quickly.  The  ad- 
dition of  rennin,  pepsin  and  hydrochloric  acid  together  caused  dur- 
ing the  first  four  hours  a  greater  digestion  of  sterilized  milk,  but  by 
longer-continued  digestion  the  unsterilized  milk  showed  a  greater 
peptone  mass.  By  the  successive  action  of  pepsin-hydrochloric  acid 
for  15  minutes,  then  rennin,  then  pancreatin  and  bile  for  4  hours, 
the  sterilized  milk  was  more  completely  digested. 

Cronheim  and  Erich  Miiller  (1903,  1908)  made  raw  and  ster- 
ilized milk  tests  with  children, — both  healthy  children  and  children 
having  rachitis.  With  the  healthy  children  no  differences  in  favor 
of  either  milk  could  be  established,  nor  were  there  marked  differen- 
ces in  the  rachitic  children. 

Other  feeding  and  metabolism  experiments  involving  the  use  of 
casein  are  those  of  Gumpert  (1905),  L.  Jacob  (1906),  Lipschiitz 
(1910a,  1911b)  and  Osborne  and  Mendel  (1911a,  1911b  and  others). 

Summary.  No  satisfactory  theory  as  to  the  production  of 
casein  in  the  milk  glands  can  be  offered  from  the  evidence  at  hand, 
nor  even  as  to  whether  the  phosphorus  for  the  same  is  obtained  di- 
rectly from  gland  nucleoprotein  or  from  the  blood. 


PHOSPHORUS  METABOLISM  275 

Artificial  digestion  tests  made  with  juices  or  enzymes  from  the 
stomach  show  that  casein  is  but  slowly  acted  on  by  such  enzymes, 
and  that  the  early  stages  of  the  process  tend  to  leave  the  main  part 
of  the  phosphorus  in  the  form  of  a  difficultly  soluble  paranuclein, 
which  may,  however,  in  time,  be  nearly  or  quite  entirely  dissolved ; 
and  that  the  phosphorus-free  products  formed  are  always  more 
readily  further  cleaved  than  are  those  containing  phosphorus,  so 
that  the  undissolved  residue  becomes  constantly  richer  in  phospho- 
rus. It  is  shown  also  that  the  phosphorus  which  goes  into  solution 
during  these  processes  is  partly  inorganic  and  partly  organic  in 
form;  and  that  the  results,  and  the  ease  and  rapidity  of  the 
action  of  pepsin  on  the  paranuclein  are  variable  for  the  casein  ob- 
tained from  different  species  of  mammalia,  the  completeness  and 
speed  of  digestion  being  greater  for  the  casein  of  woman's,  ass's 
and  mare's  milk  than  for  that  of  cow's,  goat's  and  buffalo's  milk. 
The  presence  of  carbohydrate  retards  peptic  digestion.  The  ftr- 
ment-protein  reaction  which  brings  about  cleavage,  may,  under  fav- 
oring conditions,  be  reversed. 

With  regard  to  the  enzymes  of  the  intestine,  tests  in  vitro  indi- 
cate that  activated  trypsin  may  cause  a  completion  of  the  digestion 
of  casein  as  to  both  its  nitrogen  and  its  phosphorus  portions ;  that 
the  phosphorus  thus  passing  into  solution  (very  nearly  all  of  that 
of  casein)  is  more  largely  organic  than  inorganic ;  that  the  portion 
of  the  phosphorus  converted  into  inorganic  phosphoric  acid  increases 
with  the  duration  of  digestion  and  with  the  amount  of  ferment 
present;  that  cleavage  proceeds  further  and  more  easily  under  the 
influence  of  both  pepsin  and  trypsin  than  of  either  alone ;  and  that 
bile  assists  trypsin  digestion. 

Experiments  on  living  subjects  seem  to  show  an  efficient  ab- 
sorption and  use  of  the  constituents  of  casein  by  both  animals  and 
human  beings,  older  and  younger,  perhaps  even  better  than  the  use 
made  of  flesh.  Undigested  casein,  however,  has  been  found  to  serve 
the  body  better  than  do  the  products  of  complete  pancreatin  or  acid 
cleavage  (dogs  and  mice) ,  indicating  that  within  the  organism  there 
is  some  absorption  of  casein  in  a  state  of  incomplete  digestive  cleav- 
age. Dogs  have,  however,  been  maintained  in  nitrogen  equilibrium 
on  the  products  of  complete  cleavage. 

Evidence  has  been  produced  that  of  ingested  milk  large  portions 
(four-sevenths  of  the  whole)  pass  the  pylorus  unaltered,  and  later 
the  rest  of  the  casein  passes  in  larger  or  smaller  clots,  not  pepton- 
ized. 


276  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

While  pepsin-hydrochloric  acid  digests  unsterilized  milk  more 
quickly  than  sterilized,  yet  under  the  influence  of  the  succession  of 
enzymes  present  in  the  digestive  tract,  the  sterilized  is  said  to  be 
more  completely  digested.  So  far  as  we  learn,  experiments  with 
children  have  not  demonstrated  any  practical  advantage  as  due  to* 
either  condition. 

THE  METABOLISM  OF  THE  COMPOUNDS  OF  GLYCERO- 
PHOSPHORIC  ACID 

THE  FUNCTION  OF  LECITHIN  IN  INTERMEDIARY  METABOLISM 

The  method  of  usefulness  of  lecithin  in  its  more  intimate  rela- 
tions with  the  life  of  the  animal  has  been  a  subject  of  much  spec* 
ulation,  but  little  productive  study.  A  considerable  number  of 
theories  have  been  advanced,  which  we  mention  in  brief. 

Bergell  (1898b)  concluded  that  in  animal  as  in  vegetable  cells 
organically  combined  phosphorus  stands  in  a  causal  relation  to  the 
function  of  cell  cleavage.  Lecithin  and  nuclein  phosphorus,  as  uni- 
versal cell  constituents,  may  at  least  be  regarded  as  necessary  to  cell 
cleavage. 

Loew  (1899)  suggested  that  the  chief  function  of  lecithin  is  to 
serve  for  respiration,  it  being  the  form  into  which  fat  must  be 
changed  to  become  combustible  in  the  protoplasm.  According  to 
this  theory,  by  the  transformation  of  fatty  matter  into  lecithin  the 
higher  fatty  acids  are  offered  to  the  protoplasm  in  a  soluble  form, 
and,  after  being  oxidized,  other  molecules  of  fatty  acids  may  enter 
into  the  place  of  the  former;  thus  the  same  molecules  of  glycero- 
phosphoric  acid  serving  repeatedly  as  vehicles  for  oxidation  of  mol- 
ecules of  fatty  acids. 

The  idea  of  the  service  of  lecithin  as  a  storage  product  has 
often  been  suggested.  Hanai  (1897)  considers  it  in  this  light.  He 
observed  that  old  tea  leaves  lose  their  lecithin  in  the  spring,  while 
the  amount  of  it  increases  gradually  in  the  young  leaves.  In  the 
bark  of  Prunus  cerasus  the  lecithin  also  decreases  as  the  flower  buds 
form  and  open. 

The  metabolism  of  lecithin  during  the  incubation  of  eggs,  as 
shown  by  the  work  of  Maxwell  (1893),  Mesernitzy  (1907),  Carpiaux 
(1908),  and  Plimmer  and  Scott  (1909)  (see  Incubation)  also  sug- 
gests a  storage  function;  an  accumulation  in  anticipation  of  rapid 
cell  multiplication. 

Springer  (1902)  says  that  lecithins  are  present  most  abundant- 
ly where  growth  is  most  active,  and  that  they  diminish  when  the 
seat  of  growth  shifts  to  some  other  place. 


PHOSPHORUS  METABOLISM 


277 


One  instance  in  harmony  with  this  theory  is  the  comparative 
richness  of  the  marrow  of  young  bones  in  lecithin.  Its  abundance 
in  gland  cells,  eggs,  spermatozoa  and  pollen  (Stoklasa,  1896b)  also 
suggests  a  connection  with  intense  metabolism,  but  surely  no  such 
idea  is  exemplified  by  the  high  lecithin  content  of  nerve  tissue. 

Burow  (1900)  finds  that  the  lecithin  of  the  milk  of  cattle,  dogs, 
and  human  beings  varies  directly  as  the  total  protein  of  the  milk, 
and  also  as  the  weight  of  the  brain  of  sucklings  of  the  different 
species,  in  relation  to  their  body  weight;  that  is,  the  greater  the 
relative  brain  weight,  as  compared  with  body  weight,  the  higher  is 
the  lecithin  content  of  the  milk,  reckoned  in  percent  of  the  protein. 
The  figures  are  as  follows: 


Species 

Lecithin  of  milk 
Percent 

Protein  of  milk 
Percent 

Lecithin:  Protein 
X:100 

Brain:  Body  weight 

0.054 
0.170 
0.580 

3.87 
8.05 
1.90 

1.40 
2.11 
3.05 

1-370 

Dogr 

1-30 

1:7 

Glikin  (1908a)  compared  the  lecithin  content  of  the  bodies  of 
several  species  of  young  birds  and  mammals,  and  found  that  it 
varied  directly  as  the  helplessness  at  birth.  He  suggests  an  essen- 
tial connection. 

Parrozzani  (1909)  believes  that  lecithin  represents  the  final 
stage  by  which  non-nitrogenous  organic  substances  acquire  the  pow- 
er of  combining  with  nitrogen,  especially  with  amino  acids,  for  the 
synthesis  of  albuminoid  substances. 

Hammarsten  (1905b)  suggests  a  possible  connection  between 
the  lecithin  content  of  the  bile  and  the  amount  of  fat  to  be  digested, 
and  cites  the  very  high  lecithin  content  of  the  bile  of  the  polar  bear. 

Daniel-Brunet  and  Rolland  (1911a)  suggest  the  origin  of  a  part 
of  the  biliary  lecithin  by  the  observation,  which  we  have  not  seen 
substantiated,  that  the  lecithin  of  the  bile  of  the  bull  is  decreased 
by  castration. 

As  results  of  his  studies  of  the  phosphatids  W.  Koch  has  made 
a  number  of  suggestions  as  to  the  function  of  lecithin.  In  brief 
they  are  as  follows:  In  an  early  article  (1902a)  he  advanced  the  idea 
that  the  emulsion  formed  by  the  lecithins  may  be  the  substratum 
in  which  the  reactions  of  the  cell  take  place.  The  precipitation  of 
this  emulsion  by  divalent  cations  is  prevented  by  univalent  and  tri- 
valent  cations,  as  far  as  investigated,  and  this  observation  may  fur- 
nish an  explanation  of  the  changes  brought  about  by  electrolytes  in 
the  living  cell. 


278  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Somewhat  later  (1903)  Koch  suggests  that  the  lecithins  of  the 
cell  may  have  two  values:  (1)  They,  together  with  proteins  in 
colloidal  solution,  may  furnish  the  basis  for  bringing  about  the  nec- 
essary viscosity,  by  the  facility  with  which  they  are  influenced  by 
sodium  and  calcium  ions.  (2)  They  share  in  the  metabolism  of  the 
cell,  and  especially  by  their  unsaturated  fatty  acids  and  methyl 
groups  bound  to  nitrogen,  in  causing  reactions  as  yet  not  under- 
stood. 

Koch  (1905b)  noted  the  constancy  of  the  creatinin  excretion  by 
men  and  a  dog,  and  suggested  that  the  lecithins  of  the  food  furnish 
the  methyl  groups  of  the  creatinin  excreted. 

Two  years  later  Koch  (1907a)  also  made  a  physico-chemical 
study  (colloid  precipitation  and  measurement  of  viscosity)  of  leci- 
thin and  cephalin  which  may  have  a  bearing  on  their  significance  in 
the  red  blood  cells. 

Writing  on  the  significance  of  phosphatids  for  the  living  cell, 
Koch  (1909b)  says,  — "Reactions  may  be  carried  out  with  colloidal 
solutions  of  phosphatids  which  are  very  much  like  those  observed 
with  physiological  material,  both  qualitatively  and  quantitatively. 
Both  carbonic  acid  and  ammonia  influence  lecithin  emulsion  even  in 
small  concentration,  as  was  to  be  expected  from  the  H+  and  OH-  con- 
centration, respectively.  Therefore  the  phosphatides  play  an  im- 
portant part  both  in  the  morphological  and  the  chemical  differentia- 
tion of  cells,  in  that  they  are  able  to  form  precipitation  membranes, 
which  one  may  think  of  as  distributed  throughout  the  protoplasm." 

Koch  et  al.  (1910)  *  made  a  series  of  pharmacological  studies  on 
the  phosphatids,  especially  with  reference  to  their  participation  in 
the  selection  and  transmission  of  substances  through  the  mem- 
branes of  the  cell.      Among  their  conclusions  are  the  following : 

"The  greater  concentration  of  potassium  in  the  cells  of  a  tissue 
as  compared  to  the  surrounding  lymph  spaces  or  serum  can  be  in 
part  accounted  for  by  the  specific  affinity  for  this  element  of  some 
of  the  phosphatides,  especially  kephalin." 

"There  is  no  evidence  that  anaesthetics  or  hypnotics  produce 
changes  in  the  state  of  aggregation  of  lecithin  or  kephalin,  which 
are  sufficiently  consistent  to  account  for  such  a  general  phenomenon 
as  narcosis.  There  is  some  evidence,  however,  that  chloroform,  as 
distinguished  from  pure  ether,  has  the  power  to  form  a  combination 
with  lecithin,  a  phenomenon  which  may  be  brought  into  relation 
with  its  slow  elimination  and  consequent  tendency  to  produce  de- 
layed poisoning." 

*  W.  Koch,  1910b;  Koch  and  Pike,  1910;  Koch  and  McLean,  1910;  Koch  and  Williams, 
1910;   Koch  and  Mostrom,   1910.  ' 


PHOSPHORUS  METABOLISM  279 

Considering  the  possible  participation  of  the  phosphatids, 
through  specific  chemical  affinities,  in  the  translocation  of  metab- 
olites, these  authors  reached  conclusions  a  part  of  which  are  as  fol- 
lows: 

"The  changes  in  state  of  aggregation  of  lecithin  produced  by 
sodium  chloride  are  the  result  of  the  independent  action  of  the  so- 
dium and  chlorine  ions,  whose  effects  are  in  opposite  directions.  Be- 
low the  concentration  of  a  physiological  salt  solution  (0.12  molecu- 
lar) the  action  of  the  chlorine  ion,  which  decreases  the  state  of  ag- 
gregation of  the  lecithin,  predominates.  Above  the  concentration  of 
a  physiological  salt  solution,  the  action  of  the  sodium  ion,  which 
tends  to  increase  the  state  of  aggregation  of  lecithin,  comes  more 
and  more  into  prominence. 

"It  has  been  suggested  that,  when  the  phenomenon  of  chloride 
retention  occurs,  some  change  has  taken  place  in  the  state  of  aggre- 
gation of  the  cell  lipoids  which  allows  this  action  of  the  chlorine  ion 
to  predominate  to  a  still  greater  extent. 

"Ammonia  and  bile  salts  possess  the  power  of  altering  the 
physical  state  of  aggregation  of  lecithin  to  such  an  extent  as  to  per- 
mit of  the  conclusion  that  they  can  be  of  functional  significance  in 
altering  the  permeability  of  cell  membranes. 

"The  ability  of  the  tissue  metabolites  to  combine  with  lecithin, 
as  measured  by  the  changes  in  the  physical  state  of  aggregation 
produced  by  their  presence,  is  in  some  cases  considerable,  in  other 
cases  entirely  lacking.  Thus  hypoxanthin,  creatin,  creatinin,  ad- 
renalin and  ammonia  salts  show  evidence  of  combination.  Inosite 
is  doubtful,  and  urea  is  negative. 

"The  amino  acids  show  varying  powers  of  combination.  The 
dicarboxy-acids,  like  acids  in  general,  tend  to  increase  the  state  of 
aggregation  of  lecithin. 

Studying  the  function  of  the  brain  phosphatids  in  the  action  of 
strychnin  they  conclude : 

"The  central  nervous  system,  especially  the  cord,  by  its  high 
phosphatide  content,  is  enabled  to  pick  the  strychnin  out  of  the 
blood  stream  on  account  of  the  affinity  of  the  lecithin  and  kephalin 
for  the  strychnin  as  compared  to  serum  albumin.  The  strychnin 
probably  enters  into  the  combination  with  lecithin  through  some  re- 
lation to  its  unsaturated  fatty  acid  group  (oleic  acid). 

"Strychnin  interferes  in  such  a  way  with  the  normal  relation 
of  these  unsaturated  fatty  acid  groups  to  oxygen  as  to  bring  about 
a  more  rapid  transfer  to  any  easily  oxidizable  substance." 


280  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Loewe  (1912a,  1912b,  1912c,  1912d)  made  a  series  of  physico- 
chemical  studies  of  lipoids  in  relation  to  organic  solvents  and  color- 
ing matters,  the  object  being  to  determine  the  nature  of  lipoid  ac- 
tion, biologically  and  pharmacologically. 

The  following  quotation  expresses  his  conclusion : 

"The  hypothesis  of  the  solvent  function  of  lipoids  and  the  asso- 
ciated theory  of  the  taking  up  of  substances  by  the  cells  and  of  nar- 
cosis is  not  supported  by  the  examples  studied.  The  study  of  the 
'taking  up'  by  lipoids  of  the  substances  considered  in  this  theory 
shows  that  the  process  does  not  follow  the  Henry-Nernst  absorption 
laws,  but  is  an  adsorption. 

"The  taking  up  by  lipoids  of  basic  coloring  matters,  organic 
solvents  and  organic  substances  dissolved  in  water  is  not  a  linear 
function  of  the  concentration,  but  is  determined  by  an  adsorption 
isotherm.  Therefore  it  is  not  to  be  expected  that  the  lipoid  com- 
ponents of  the  cell  membrane,  so  far  as  they  are  not  very  freely 
movable  on  the  cell  contents,  will  serve  to  facilitate  the  taking  up 
into  the  interior,  but  rather  their  action  will  be  hysteresis." 

Reicher  (1911)  believes  that  the  physiologic  oxidation  of  fat, 
in  general,  is  made  possible  only  through  combination  in  lecithin,  an 
idea  in  harmony  with  that  of  Loew  (1899).  The  feeding  of  leci- 
thin-free fats,  and  the  perfusion  of  a  surviving  liver  with  blood  and 
triolein  increased  the  lecithin  of  the  blood. 

Kovaliova  (1912)  compared  lecithin  and  other  phosphorus  com- 
pounds with  reference  to  their  effects  on  the  power  of  oxidation  in 
rabbits.  The  phosphorus  compounds  were  administered  by  subcu- 
taneous injection  in  oil  suspension.  Two  hours  later  the  rabbits 
were  injected  with  2  gm.  benzene,  and  placed  in  an  apparatus  where 
the  respiration  coefficient  and  power  of  oxidation  were  determined. 
Five-hour  and  24-hour  experiments  were  performed. 

Lecithin  caused  an  increase  in  the  elimination  of  phenol,  and  in- 
crease in  the  respiration  coefficient.  Sodium  nucleate  in  small 
doses  caused  an  elevation,  but  in  large  doses  a  depression,  of  respira- 
tion coefficient  and  degree  of  oxidation.  Sodium  glycerophosphate 
caused  a  depression  of  the  respiration  coefficient,  but  a  greater  in- 
crease of  the  degree  of  oxidation  than  that  of  the  two  preceding  sub- 
stances. Phytin  in  aqueous  suspension  caused  an  increase  in  the 
degree  of  oxidation,  but  scarcely  affected  the  respiration  coefficient. 
In  Kovaliova's  judgment  lecithin  exerted  the  more  uniform  influ- 
ence on  the  elevation  of  the  respiration  coefficient  and  degree  of 
oxidation,  while  the  sodium  nucleate,  in  large  doses,  exerted  the 
most  uniform  action  in  lowering  them. 


PHOSPHORUS  METABOLISM  281 

Stuber  (1913a,  b)  determined  the  effect  of  lecithin  and  choles- 
terol on  the  phagocytic  index  of  leucocytes.  Cholesterol  diminishes 
the  index.  Lecithin  does  not  affect  it,  but  prevents  the  action  of 
cholesterol.  The  oleic  and  palmitic  acid  esters  of  cholesterol  also 
check  phagocytosis,  but  lecithin  does  not  prevent  this  action. 

Maslow  (1913)  studied  the  effects  of  lecithin  and  other  phos- 
phorus compounds  on  the  functions  of  the  intracellular  ferments  of 
dogs.  Several  litters  of  young  dogs  were  the  subjects  of  these  ex- 
periments. One  was  used  as  a  control,  being  killed  at  the  begin- 
ning of  the  experiment;  and  a  second  control  was  fed  the  usual 
mixed  meat  diet,  and  was  then  killed  before  the  close  of  the  ex- 
periment. The  remaining  animals  were  divided  into  four 
groups.  One  received  a  low-phosphorus  broth  containing  pro- 
tein, carbohydrate  and  fat  in  sufficient  quantity ;  another  group  re- 
ceived the  same  broth  plus  a  phosphate;  a  third,  the  same  plus 
glycerophosphate;  and  a  fourth,  lecithin.  The  second  group  re- 
ceived either  whole  milk,  or  a  milk  preparation  in  which  the  casein 
was  replaced  by  albumin. 

Under  normal  conditions  of  development  there  was  in  all  organs 
a  normal  development  of  fermentative  energy.  On  the  phosphorus- 
poor  ration  the  animal  declined  in  weight,  and  died.  The  phos- 
phorus content  of  the  organs  decreased,  the  decrease  being  mostly 
in  the  inorganic  phosphorus.  Of  the  organic  phosphorus  only  the 
lipoid  phosphorus  decreased.  The  ferment  function  of  the  organs 
was  markedly  disturbed ;  the  catalase,  lipase,  amylase,  diastase,  and 
nuclease  were  depressed  in  amount  or  checked  in  their  development. 

The  addition  of  inorganic  and  glycerophosphate  phosphorus  to 
the  diet  were  without  avail.  The  organs  showed  phosphorus  im- 
poverishment and  the  intracellular  enzymes  were  not  favorably  in- 
fluenced. Lecithin,  however,  increased  the  phosphorus  content  of 
the  organs,  especially  their  organic  phosphorus  content,  and  stimu- 
lated the  development  of  the  ferments. 

The  author  concludes  that  abundance  of  assimilable  phosphorus 
in  the  food  runs  parallel  with  abundant  ferment  formation  in  the  or- 
ganism, and  that  a  power  of  the  animal  to  synthesize  organic  phos- 
phorus compounds  is  not  probable.  For  therapy  the  conclusion 
was  drawn  that  the  use  of  phosphates  and  glycerophosphates  should 
be  limited,  and  that  lecithin  is  to  be  preferred,  if  increase  of  the 
phosphorus  content  of  the  organism  or  activating  of  the  ferments 
is  desired. 

Mayer  and  Schaeffer  (1913)  find  that  the  lipoid  content  of  tis- 
sues tends  strongly  to  remain  characteristic  of  the  species  and  tis- 
sue, neither  fasting  nor  overfeeding  producing  characteristic 
change,  this  proportion  therefore,  appearing  to  be  fundamental  and 


282  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

permanent.  The  ratio  of  fatty  acids  to  lipoid  phosphorus  is  re- 
markably constant  in  certain  types  of  cells  in  different  animals,  as 
for  instance  in  kidney  and  red  blood  cells.  A  relation  is  shown  to 
exist  between  the  capacity  of  tissues  to  imbibe  water,  and  the  ratio 
of  lipoid  phosphorus  to  cholesterin.  In  all  the  species  examined  the 
various  organs  compared  as  to  lipoid  phosphorus  content  (on  the 
fresh  basis)  in  the  same  order;  their  content  of  lipoids  apparently 
being  related  to  physiologic  activity,  in  ways  yet  to  be  explained. 

Lecithin  and  other  phosphatids  of  the  blood  corpuscles  and  of 
the  tissue  cells  also  stand  in  important  relations  to  the  action  of 
poisons  of  various  sorts  and  origin,  an  important  subject  which  has 
not  been  included  in  this  investigation. 

THE  INFLUENCE  OF  LECITHIN  IN  DIGESTION 

There  is  prevalent  an  idea  that  lecithin  is  of  functional  value 
in  the  alimentary  digestion  processes,  but  the  experimental  evidence 
on  the  subject  is  not  all  in  support  of  the  theory.  Thus  Kalabou- 
koff  and  Terroine  (1907a,  1907b,  1907c)  tested  lecithin  in  relation  to 
the  cleavage  of  monobutyrin  and  olive  oil ;  also  as  to  its  influence  in 
activating  pancreatic  juice  to  such  cleavage,  with  and  without  the 
presence  of  bile  salts.  Such  activation  by  bile  salts  was  quite  evi- 
dent, whether  with  or  without  lecithin ;  but  lecithin  showed  no 
such  activating  power  in  the  case  of  monobutyrin,  and  only  a  slight 
influence,  and  at  relatively  high  concentration  in  the  case  of  olive 
oil.  This  conclusion  is  in  harmony  with  those  of  0.  vonFiirth  and 
Jul.  Schulz. 

In  the  second  paper  were  reported  similar  tests  of  the  activat- 
ing power  of  lecithin,  and  of  bile  salts  with  gastric  and  intestinal 
juices.  "1.  The  lipase  of  the  glycerin  extract  of  gastric  mucous  is 
not  at  all  changed  by  the  addition  of  lecithin ;  it  is  notably  retarded 
by  bile  salts.  2.  The  intestinal  lipase  is  not  at  all  modified  by  the 
addition  of  lecithin;  it  is  activated  by  bile  salts." 

The  third  paper  had  to  do  with  the  action  of  ovo-lecithin  on 
amylase,  trypsin  and  rennet.  "Addition  of  lecithin  does  not  modify 
saccharification  of  starch,  the  digestion  of  casein  and  coagulated  al- 
bumin, nor  the  coagulation  of  milk  by  pancreatic  juice;  the  addition 
of  bile  salts  activates  plainly  the  coagulation  of  milk  by  kinased  pan- 
creatic juice." 

In  a  later  paper  (Kalaboukoff  and  Terroine,  1909)  cleavage  of 
an  emulsion  of  pure  lecithin  (prepared  from  egg  yolk)  by  pancre- 
atic juice  at  40°  was  tested  by  the  acidity  developed.  No  increase 
of  acidity  was  detected  when  boiled  pancreatic  juice  was  used,  and 
but  slight  increase  when  unboiled  juice  was  used;  this  was  a  little 
further  increased  by  the  presence  of  bile  salts;  but  even  after 
66  hours  the  acidity  was  but  slight  (not  more  than  equivalent  to 


PHOSPHORUS  METABOLISM 


283 


2.8  c.c.  of  N/20  NaOH  for  10  c.c.  of  the  mixture).  These  authors 
think  that  the  results  found  by  Stassano  and  Billon  (1903a,  1903b) 
should  be  referred  to  cleavage  of  fatty  substances  obtained  in  the 
course  of  preparation. 

Regarding  some  other  phases  of  this  matter  these  authors  say : 
"The  work  of  Slowtzoff  and  of  Stassano  and  Billon  has  shown  that 
lecithin  is  absorbed  and  appears  in  lymph.  Bayer  has  found,  and 
so  do  we,  that  if  a  solution  of  bile  salts  be  added  to  a  milky  lecithin 
emulsion  a  clear  liquid  results.  Ultramicroscopically,  lecithin  emul- 
sions have  a  colloidal  appearance,  with  an  infinite  number  of  grains, 
and  these  almost  disappear  after  the  addition  of  a  solution  of  bile 
salts." 

As  a  result  of  recent  investigations,  Terroine  (1911)  repeats 
his  conclusions :  (1)  Lecithin  does  not  increase  the  rate  of  hydroly- 
sis of  monobutyrin  by  pancreatic  juice,  and  increases  only  slightly 
in  more  concentrated  solution  the  rate  of  hydrolysis  of  oils.  (2) 
It  increases  the  lipolytic  action  of  neither  mucous  membrane  of 
stomach  (in  glycerol)  nor  intestinal  lipase.  (3)  It  has  no  action 
on  the  rate  of  hydrolysis  of  starch,  the  digestion  of  milk  casein  or 
coagulated  albumin,  or  the  coagulation  of  milk  by  pancreatic  juice. 

On  the  other  side  of  this  question  we  have  the  work  of  Moore 
and  Parker,  Hewlett,  Loevenhart  and  Souder,  Kiittner  and  Usuki  as 
mentioned  in  brief  below. 

Moore  and  Parker  (1901)  state  that  the  presence  of  lecithin 
greatly  increases  the  solvent  power  of  bile  toward  fatty  acids  and 
soaps.  The  bile  salts  serve  to  keep  lecithin  and  cholesterin  in  solu- 
tion, and  hence  aid  in  their  emulsification  by  the  liver.  The  com- 
bination of  bile  salts  and  lecithin  accomplishes  the  solution  of  the 
fatty  acids  and  soaps.  Numerical  results  from  this  study  are  given 
below. 

SOLUBILITY  OF  FATS  AND  SOAPS  AS  AFFECTED  BY  BILE  SALTS  AND 

LECITHIN 


Distilled  water 

Bile  salts, 
5  percent 

Bile  salts,  5  percent 
+lecithin,  1  percent 

Practically  insoluble 
Less  than  0t.l*  soluble 

0.5*  soluble 
0.5 
0.1 
Less  than  0. 1% 

0.7*  soluble 

4. 

0.6 

0.2 

2.2396                   ) 
apparently  colloid  sol.  f 
5.0*  soluble 
0.2 
0.1 
(Less  than  0.1*) 
(Much  less  than  0.1%) 

Not  tried 

7.6* 
1.0 
0.2 
0.2 
Less  than  0. 1* 

Not  tried 

11.6% 
2.4 
0.7 
1.4 
0.9 
0  4 

(Less  than  0.1*) 
(Much  less  than  0#.  1*) 

Practically  insoluble 
Absolutely  insoluble 

3.2 

0.2 

Less  than  0.1* 

7.0* 
About  0.1* 

8.2 
1.2 
1.0 

Not  more  than  0. 15% 

Free  fatty  acids 

mixed., 

oleic 

palmitic 

stearic 

Sodium  salts 

mixed 

oleate , 

palmitate 

stearate. 

Calcium  salts,  oleate .... 

palmitate 

stearate 

Magnesium  salts,  oleate 

palmitate 

stearate 

Lecithin 

Cholesterin 


284  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Reiss  (1904)  determined  that  lecithin,  with  chloroform,  is  able 
to  take  up  the  ferments  rennet  and  trypsin,  as  neither  constituent 
alone  can,  and  as  no  other  fat  tried  can  with  chloroform.  It  is 
thought  that  this  may  have  some  significance  with  ref- 
erence to  the  role  of  lecithin  in  the  body.  This  is  likened  to 
the  activating  power  of  lecithin  toward  the  haemolysis  of  cobra 
poison. 

Hewlett  (1905)  studied  the  action  of  lecithin  in  bile  on  fat  di- 
gestion by  pancreatic  juice  of  a  dog.  Bile  increased  the  action  of 
pure  pancreatic  juice  on  triacetin.  (Fat  was  not  used,  to  eliminate 
the  factor  of  influence  through  emulsification.  Triacetin  is  a  sol- 
uble glyceride.)  Bile  served  to  increase  the  action  of  pure  pancre- 
atic juice  on  this  ester,  especially  in  the  first  hour.  Boiling  the 
bile  does  not  destroy  this  property,  and  furthermore,  it  was  found 
not  to  reside  in  cholesterin  or  the  pigments,  and  was  found  not  to 
depend  on  variations  in  reaction,  nor  on  variation  in  the  amount 
of  calcium  salts  present.  Precisely  the  same  effects  of  accelera- 
tion were  produced,  however,  by  the  addition  of  lecithin  to  the  pan- 
creatic juice.  Hewlett  suggests  that  this  influence  is  of  the  nature 
of  a  "zymoexcitor." 

Loevenhart  and  Souder  (1906-7)  investigated  the  relations  of 
lecithin  to  the  digestion  of  the  higher  fats  by  quantitative  determin- 
ations of  the  activity  of  pancreatic  juice  on  olive  oil,  ethyl  butyrate, 
diacetin,  triacetin,  ethyl  acetate  and  ethyl  propionate  in  the  pres- 
ence and  absence  of  bile  salts,  and  bile. 

Bile  salts,  lecithin  and  bile  greatly  accelerate  the  action  of  pan- 
creatic juice  on  all  the  esters  studied.  The  effect  of  these  accel- 
erators differs  with  the  particular  ester,  and  differences  of  experi- 
mental conditions  greatly  alter  the  degree  of  acceleration,  and  the 
relative  activity  of  the  bile  salts  and  lecithin.  These  accelerating 
capacities  are  considered  to  depend  to  a  certain  extent  on  their  sol- 
vent action,  and  also  on  their  action  on  the  enzyme  in  some  other 
way. 

Kiittner  (1907)  determined,  in  vitro,  the  effect  of  lecithin  on  the 
peptic  digestion  of  casein,  and  on  pancreatic  digestion  of  monobutyl 
glycerin.  Lecithin  in  certain  proportions  accelerates  the  enzyme 
action  of  gastric  or  pancreatic  juice ;  in  other  greater  proportions  it 
retards  it.  Curves  were  plotted  exhibiting  this  action.  The  cause 
of  this  action  was  not  determined. 

Long  and  Gephart  (1908b)  report  that  bile  salts,  as  ordinarily 
obtained,  carry  down  a  phosphorus  complex,  which,  by  Hammarsten 
and  others,  is  regarded  as  lecithin,  and  also  that  bile  salts  are  capa- 
ble of  dissolving  and  holding  80  percent  of  their  weight  of  egg 
lecithin. 


PHOSPHORUS  METABOLISM  285 

Usuki  (1910)  studied  the  influence  of  lecithin  on  fat  digestion 
in  the  stomach,  and  in  the  intestine,  by  the  feeding  of  dogs;  and 
after  certain  lengths  of  time  the  dogs  were  killed,  the  different 
portions  of  the  alimentary  tract  separated  by  tying,  and  the  con- 
tents removed  and  analyzed.  The  results  seem  unmistakable.  The 
author's  conclusions  are  as  below. 

1.  According  to  these  experiments  the  digestion  of  fat  took 
place  more  slowly  after  feeding  of  milk  than  after  feeding  of  milk- 
lecithin  or  milk-yolk  mixtures  which  left  the  stomach  l1/^  to  2  hours 
sooner  than  the  milk  alone.  While  of  the  pure  milk,  after  six 
hours,  only  half  had  left  the  stomach,  the  same  result  was  obtained 
with  the  mixtures  in  four  hours. 

2.  After  taking  milk  more  soaps  are  found  in  the  contents  of 
the  small  intestine  than  after  the  taking  of  milk-lecithin  mixture. 
On  the  other  hand,  after  taking  milk-yolk  mixture,  more  soaps  were 
found  in  the  feces.  This  contradictory  relation  between  the  con- 
tents of  the  small  intestine  and  the  feces  is  probably  to  be  ex- 
plained by  absorption  in  the  large  intestine. 

3.  Splitting  of  lecithin  in  the  stomach  takes  place  more  quick- 
ly than  splitting  of  neutral  fat. 

'  4.  Lecithin  acts  favorably  on  the  saponification  of  the  neutral 
fat.  This  is  to  be  explained  by  the  fact  that  the  digestion  of  fat 
is  accelerated  by  the  presence  of  lecithin. 

5.  In  spite  of  the  lower  lecithin  content  of  the  milk-yolk  mix- 
ture the  digestion  of  fat  after  taking  said  mixture  was  just  as  good 
as  after  the  milk-lecithin  mixture.  The  egg-yolk  has  this  advan- 
tage because  of  the  fineness  of  the  emulsion  and  the  chemical  nature 
of  fat  contained  in  it. 

6.  It  has  been  discovered  that  the  passing  of  the  milk  fat 
from  the  stomach  did  not  begin  until  the  percentage  content  of  fat- 
ty acids  had  about  reached  the  maximum  (2  hours  after  feeding). 
This  percent  of  fatty  acids  was  maintained  almost  unchanged  dur- 
ing the  further  course  of  the  digestion.  Only  during  the  last 
stages  of  the  digestion  does  the  percent  of  fatty  acids  rise  again. 

7.  No  lecithin  was  found  in  the  intestine.  It  must,  therefore, 
have  been  split  either  in  the  stomach  or  immediately  after  entrance 
into  the  intestine. 

Lapidus  (1910)  announced  results  of  preliminary  experiments 
on  the  effect  of  lecithin  on  animal  diastases.  Lecithin  in  certain  con- 
centrations affected  unfavorably  the  activity  of  salivary,  pancreatic, 
serum  and  intestinal  diastases  in  water  and  glycerin  extracts,  but 
activated  serum  diastase  in  ether  solution. 


286  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Bickel  (1911)  suggests  an  indirect  influence  of  the  lecithin, 
leading  to  the  better  digestion  and  use  of  phosphorus  from  other 
parts  of  the  diet. 

Minami  (1912)  found  that  lipoids  are  superfluous  for  diastatic 
action  by  saliva. 

Summary.  As  a  result  of  these  investigations  we  conclude 
that  under  certain  experimental  conditions,  at  least,  lecithin  may 
assist  not  only  in  the  solution  of  fats  and  soaps,  but  also  may  accel- 
erate the  pancreatic  cleavage  of  fats,  and  even  the  peptic  digestion 
of  casein.  These  actions  vary  much  with  the  particular  nutrient 
compounds  involved,  and  with  the  experimental  conditions,  especi- 
ally as  regards  concentration  of  the  lecithin  solution.  At  certain 
concentrations  there  may  be  acceleration  of  digestion,  and  at  others 
a  retarding  action. 

In  the  actual  digestion  of  food  within  the  alimentary  tract  leci- 
thin seems  to  accelerate  the  digestion  of  fat,  and  also  the  passage  of 
food  from  the  stomach. 

DIGESTION  OF  LECITHIN  AND  GLYCEROPHOSPHATES 

That  the  digestion  of  lecithin  may  involve  its  cleavage  has  been 
observed  by  a  considerable  number  of  workers,  among  the  earlier 
ones  being  Bokay  (1877),  who  concluded  from  artificial  digestion 
experiments  and  from  feeding  experiments  with  dogs,  that  lecithin 
is  split  in  digestion  by  the  fat-splitting  ferment  of  the  pancreas, 
or  by  putrefactive  ferments  in  the  intestine;  and  Hasebroek  (1888), 
who  concluded,  from  fermentation  experiments,  that  the  phospho- 
rus from  lecithin  must  be  absorbed  as  glycerophosphoric  acid ;  also 
Nesbitt  (1899a,  1899b),  who  demonstrated  the  presence  in  the  in- 
testine of  the  dog  of  choline,  neurine  and  an  unidentified  ptomaine 
after  the  ingestion  of  egg  yolk  and  the  occlusion  of  the  small  in- 
testine at  its  lower  end.  See  also  Gaston  (1902).  Among  the 
later  workers  to  demonstrate  the  digestive  cleavage  of  lecithin  are 
Pvogozinski  (1910),  Usuki  (1910),  and  Yoshimoto  (1910). 

The  cleavage  products,  as  a  rule,  are  choline,  fatty  acids  and 
glycerophosphoric  acid,  but,  according  to  J.  and  W.  Cronheim  (1911), 
the  decomposition  may  go  only  to  distearyl-phosphoric  acid.  The 
phosphorus  of  lecithin,  then,  is  absorbed,  in  the  main,  in  the  same 
form  as  though  ingested  as  glycerophosphate.  That  a  portion  of  the 
lecithin  is  absorbed  without  cleavage,  however,  has  been  demon- 
strated by  Slowtzoff,  and  others,  as  will  be  mentioned  at  greater 
length  below. 


PHOSPHORUS  METABOLISM  287 

Marfori  (1905)  studied  the  elimination  of  glycerophosphoric 
acid  ingested  by  dogs  and  men.  In  men  it  was  found,  if  adminis- 
tered in  suitable  amounts,  to  be  absorbed  from  the  alimentary 
canal.  In  dogs  it  is  largely,  though  not  completely,  absorbed,  a 
part  being  decomposed  in  the  alimentary  tract.  At  all  events  it  is 
an  easily  assimilable  compound. 

Paul  Mayer  (1905)  finds  that  the  lipase  of  intestinal  juice  splits 
lecithin  (racemic)  asymmetrically;  the  dextro-lecithin  is  then  de- 
composed, and  the  laevo-lecithin  is  not.  From  artificial  digestion 
tests  of  lecithin  with  lipase  (1906b),  and  from  a  study  of  the  race- 
mic and  the  laevo-  forms  as  to  optical  activity,  structure  and  cleav- 
age products,  the  author  concludes  as  reported  above.  The  break- 
down of  the  dextro-lecithin  results  in  fatty  acids  and  dextro-glycero- 
phosphoric  acid.  It  is  suggested  that  the  asymmetric  cleavage, 
and  difference  in  digestibility  of  the  fractions,  may  have  practical 
significance ;  since  the  various  lecithin  products  used  medicinally  do 
differ  in  optical  activity,  that  is,  are  more  or  less  strongly  racemized, 
it  cannot  be  thought  surprising  that  the  results  of  lecithin  therapy 
are  very  unequal,  considering  the  different  reactions  of  the  enzymes 
on  dextro-  and  laevo-lecithin. 

Kutscher  and  Lohmann  (1903)  found  that  the  ferment  of  the 
pancreas  caused  lecithin  to  break  down  into  glycerophosphoric  acid, 
fatty  acids  and  neurine.  Very  little  lecithin  was  decomposed  by 
the  gastric  enzyme. 

Stassano  and  Billon  (1903a)  concluded  from  microscopic  ex- 
amination that  pancreatic  juice  does  not  cleave  lecithin,  and  after 
feeding  pure  lecithin  and  egg  yolk  (1903b),  and  then  examining  the 
chyle  with  a  polarizing  microscope,  they  concluded  that  leci- 
thin, taken  pure, — not  in  albuminoid  combination — escapes  the 
digestive  juices;  and  enters  the  circulation  by  way  of  the  chyle 
unaltered. 

Coriat  (1904a)  found  that  neither  trypsin  nor  pepsin  can  split 
lecithin,  but  that  lipase  is  able  to  do  so.  Pepsin  and  trypsin  seem 
even  to  inhibit  the  normal  enzymatic  autolysis  of  brain  tissue 
through  which  choline  is  liberated.  Coriat  finds  that  this  autolytic 
enzyme  acts  only  in  neutral  or  slightly  alkaline  media,  that  the  pro- 
duction of  choline  is  greater  in  the  latter  than  in  the  former,  that 
the  enzyme  is  inactive  in  slightly  acid  media,  and  that  this  enzyme 
can  be  destroyed  by  heat. 

Slowtzoff  (1906a)  finds  that  lecithin  given  per  os  acts  the  same 
as  lecithin  given  hypodermically,  but  that  more  must  be  given  to 
produce  the  same  result. 

In  eight  experiments  dogs  were  given  egg  yolk  or  lecithin,  and 
the  lymph  was  collected  for  analysis  after  4-5  hours ;  also  in  four 


288  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

experiments  the  intestinal  contents  of  a  dog,  after  it  had  ingested 
lecithin,  were  analyzed  for  lecithin  and  its  splitting  products ;  fur- 
ther, digestion  experiments  were  conducted  with  lecithalbumin,  and 
with  digestive  ferments  in  vitro. 

Slowtzoff  determined  that  lecithin  in  appreciable  quantities 
reaches  the  blood  unchanged,  as  well  as  in  a  saponified  condition,  by- 
way of  the  chyle,  but  not  by  the  portal  vein ;  that  lecithin  may  be 
saponified  by  the  pancreatic  juice,  or  even  by  steapsin;  that  the 
cleavage  of  lecithin  to  glycerophosphoric  acid  and  choline  occurs 
only  in  the  duodenum ;  that  pepsin-hydrochloric  acid  digestion  does 
not  affect  lecithin,  but  that  lecithalbumins  are  digested,  the  acid- 
albumin  first  formed  being  combined  with  the  lecithin,  in  which 
form  lecithin  may  perhaps  be  absorbed;  the  albumose  solution 
resulting  from  pepsin  digestion  being  free  from  lecithin.  This  lecith- 
acid  albumin,  when  introduced  per  rectum,  disappears. 

Long  (1906b)  finds  ether-soluble  phosphorus  in  human  feces 
equivalent  to  1-5.5  gm.  of  lecithin  per  day.  He  quotes  Deucher  as 
finding  in  the  feces  of  a  man  with  closed  pancreatic  duct  as  high  as 
8  grams  of  lecithin  per  day.  Long  and  Johnson  (1906)  suggest 
that  feces  lecithins  may  be  but  remotely  related  to  simple  distearyl 
lecithin,  and  also  suggest  that  the  feces  lecithin  may  come  in  part 
each  from  food,  intestinal  epithelium,  bile  residues  and  bacteria. 

Schumoff-Simanowski  and  Sieber  (1906)  state  that  lecithin 
(from  egg  yolk,  and  Kahlbaum's)  is  split  by  steapsin  from  the  pan- 
creas and  from  the  stomach,  more  energetically  by  the  former. 
Plant  ferments,  in  particular  that  from  the  seeds  of  castor-oil  plant, 
break  it  up  also,  and  in  the  same  way,  by  splitting  off  fatty  acids. 
The  lipase  of  blood  of  different  animals  does  not  cleave  it. 

Hamill  (1906-7)  determined  by  collection  and  analysis  of  chyle 
from  a  fistula  in  the  thigh  that  lecithin  administered  by  the  mouth 
produced  a  rise  in  the  ether-soluble  phosphorus  of  the  chyle. 

Franchini  (1907,1908a)  sought  to  determine  the  fate  of  that 
portion  of  ingested  lecithin  which  is  absorbed  without  cleavage.  By 
analysis  of  the  bodies  of  rabbits  after  the  feeding  of  lecithin  he 
found  a  distinct  rise  of  the  lecithin  content  of  both  liver  and  muscle, 
but  not  of  brain,  following  the  ingestion  of  lecithin  either  during 
feeding  or  fast.  There  was  also  perhaps  a  slight  increase  of  leci- 
thin phosphorus  in  the  excreta.  No  choline  was  found  in  the  urine, 
but  formic  acid  from  the  cleavage  and  oxidation  of  choline  is  said  to 
have  been  present. 

Marfori  (1908b)  in  discussing  the  assimilation  of  organic  phos- 
phorus compounds  says: 


PHOSPHORUS  METABOLISM  289 

"Phosphorus  in  the  form  of  synthetic  glycerophosphoric  acid, 
taken  into  the  body  by  the  mouth,  is  easily  absorbed  and  assimilated. 
Lecithin  administered  either  per  os  or  subcutaneously  is  taken  up  at 
once  by  the  tissues  for  their  growth." 

Clementi  (1910)  finds  that  pancreatic  juice  hydrolyzes  the  fatty 
acid  radicals  in  lecithin;  an  action,  however,  which  is  not  equally 
noticeable  with  all  pancreatic  juices,  but  which  varies,  in  a  more  or 
less  pronounced  way,  according-  to  the  capacity  of  the  juices  to 
saponify  the  ordinary  glycerides.  Intestinal  juice  lipase,  as  well  as 
pancreatic  juice  lipase,  acts  on  lecithin. 

Mathison  (1910)  found  synthetic  glycerophosphoric  acid  not  de- 
composed by  pepsin,  or  trypsin,  or  by  fresh  pancreatic  juice,  either 
with  or  without  enterokinase.  He  thinks,  therefore,  that  glycero- 
phosphoric acid  is  absorbed  as  such,  though  he  notes  the  possibility 
that  the  natural  and  the  artificial  products  may  behave  differently 
in  this  matter. 

Brugsch  and  Masuda  (1911)  report  that  the  intestinal  juice 
contains  a  lecithin-cleaving  ferment,  and  also  that  extracts  of  the 
colon  bacillus  and  Staphylococcus  cleave  lecithin. 

Grosser  and  Husler  (1912)  determined  that  the  mucous  mem- 
brane of  the  intestine,  and  the  cells  of  the  kidney,  contain  a  ferment 
which  completely  splits  glycerophosphate  solutions.  The  lungs  al- 
so contain  such  a  ferment,  but  apparently  in  smaller  amount.  The 
liver  and  spleen  show  its  presence  but  sparingly,  while  pancreas, 
muscle,  heart  muscle  and  blood  are  free  from  such  a  ferment.  The 
feces  contain  this  enzyme  but  the  urine  does  not.  Sodium  glycero- 
phosphate and  the  glycerophosphoric  acid  from  natural  lecithin  were 
alike  decomposed  by  this  enzyme. 

Ehrmann  and  Kruspe  (1913)  determined  that  the  exclusion  of 
bile  from  the  alimentary  tract  greatly  lowers  the  absorption  of  leci- 
thin or  causes  its  elimination  in  considerable  quantity  in  the  feces ; 
and  the  exclusion  of  the  pancreatic  secretion  produces  a  similar 
though  less  pronounced  effect.  These  authors  conclude  that  the 
total  lecithin  is  not  split  by  the  pancreatic  juice,  but  that  a  portion 
is  absorbed  as  such.  See  also  Forbes  and  associates  (1914)  under 
Nutr.  Val.  Org.  and  Inorg.  P. 

Summary.  From  the  above  investigations  we  conclude  that 
lecithin  is  absorbed  in  part  without  digestive  cleavage,  in  part  after 
separation  into  choline,  fatty  acids  and  glycerophosphoric  acid, 
probably  in  part  combined  as  lecith-acid  albumin,  and  probably  also 
in  part  after  the  splitting  of  the  glycerophosphoric  acid  to  glycerin 
and  inorganic  phosphoric  acid. 


290  OHIO  EXPEKIMENT  STATION:  TECHNICAL  BUL.  5 

The  digestive  cleavage  of  lecithin  is  accomplished  by  the  fat- 
splitting  enzymes  of  the  digestive  tract,  mostly  in  the  duodenum. 

Lecithin,  as  such,  reaches  the  circulation  by  way  of  the  chyle, 
but  not  by  the  portal  vein. 

The  feces  lecithin  probably  comes  in  part  each  from  food,  in- 
testinal epithelium,  bile  residues  and  bacteria. 

Lecithin  taken  by  the  mouth,  or  hypodermically,  may  be  assim- 
ilated at  once  by  the  tissues. 

Glycerophosphates  are  absorbed  in  part  as  such,  and  in  part 
after  cleavage  to  glycerin  and  inorganic  phosphates. 

BALANCE  EXPERIMENTS  WITH  LECITHIN  AND  OTHER  COMPOUNDS 
OF  GLYCEROPHOSPHORIC  ACID 

Of  the  many  observations  on  the  nutritive  value  of  compounds 
of  glycerophosphoric  acid  comparatively  few  are  of  the  nature  of 
complete  balance  experiments.  Additional  evidence  of  this  sort, 
especially  comparing  these  with  other  phosphorus  compounds, 
would  be  of  value. 

That  the  phosphorus  of  glycerophosphates  is  absorbed  and  re- 
tained is  universally  admitted.  An  experiment  demonstrating  this 
point  is  that  of  Sanson  (1896),  who  fed  calcium  glycerophosphate  to 
rabbits. 

Considerable  interest  has  attached  to  the  function  of  lecithin 
in  infant  nutrition,  especially  as  added  to  the  milk  in  the  form  of 
egg  yolk.  Cronheim  and  Miiller  (1900)  compared  egg  yolk  and 
milk  powder  as  elements  in  the  diet  of  a  child  eleven  and  a  half 
months  old.  The  ration  containing  the  egg  yolk  was  rich  in  leci- 
thin ;  the  other  was  poor  in  lecithin.  The  calorific  value  of  the  lat- 
ter was  a  trifle  higher  than  that  of  the  former.  Nitrogen  and 
phosphorus  retention,  and  gain  in  weight  all  showed  the  ration  con- 
taining the  egg  yolk  to  be  the  more  efficient. 

In  a  later  series  of  experiments  Cronheim  and  Miiller  (1902)  made 
further  comparisons  of  protein  phosphorus  and  lecithin  phosphorus 
in  metabolism  experiments  with  children,  dogs  and  guinea  pigs. 
The  periods  with  children  averaged  four  days  in  length,  which  is  in- 
sufficient for  mineral  balance  experiments.  The  results  on  infants 
are  inconclusive. 

Five  young  dogs  were  fed  on  similar  rations  to  those  received 
by  the  children.  Egg  yolk  was  compared  with  plasmon — a  milk  al- 
bumin preparation.  These  dogs  were  fed  for  3  months,  after  which 
they  were  killed,  and  the  bones  subjected  to  study.  The  egg  yolk 
had  produced  better  bone  development,  and  the  marrow  produced 


PHOSPHORUS  METABOLISM 


291 


by  the  ration  containing  this  food  was  yellow  and  rich  in  fat,  while 
that  from  the -bones  of  the  dogs  which  had  received  plasmon  was 
red  and  immature. 

Egg  yolk  and  plasmon  were  also  compared,  using  guinea  pigs 
as  subjects.  The  animals  receiving  the  egg  yolk  grew  the  more 
rapidly  and  had  the  fatter  livers. 

Cronheim  and  Muller  conclude  that  egg  yolk,  apparently 
through  its  lecithin  content,  especially  favors  nitrogen  retention, 
and  is  to  be  recommended  for  use  in  the  early  feeding  of  the  child. 

Zuntz  (1900)  also  conducted  an  experiment  in  infant  metabo- 
lism in  which  a  child,  eleven  and  a  half  months  old,  was  fed  for  two 
days  on  a  milk  diet  in  which  6  percent  of  the  dry  matter  was  replaced 
by  egg  yolk,  followed  by  a  period  of  the  same  length  on  milk  alone. 
While  receiving  the  egg  yolk  the  child  retained  24.2  percent  of  the 
nitrogen  of  the  ration;  on  the  milk  diet  the  nitrogen  retention 
was  9.9  percent  of  the  intake ;  of  the  phosphorus  of  the  egg  ration 
33.3  percent  was  retained,  and  of  the  diet  of  milk  alone  17.4  percent 
was  retained. 

Lebbin  (1901)  conducted  a  2-day  metabolism  experiment  on  a 
man  28  years  old,  and  weighing  65  kg.,  using  eggs,  of  which  22  were 
consumed,  as  the  only  food.  The  intake  contained  286.28  gm.  dry 
matter,  and  the  outgo  14.28  gm.  The  eggs  contained  39.22  gm.  leci- 
thin, and  the  excreta  3.517  gm. 

Massacin  (1902)  conducted  a  series  of  balance  experiments 
with  a  man  suffering  from  pulmonary  catarrh,  on  a  normal  mixed 
diet  with  lecithin  added  in  certain  periods.  The  addition  of  lecithin 
seems  to  have  increased  nitrogen  retention. 

Gilbert  and  Posternak  (1903)  conducted  a  balance  experiment 
on  a  man,  for  the  purpose  of  learning  the  effect  of  lecithin  ingestion 
on  phosphorus  metabolism.      The  results  are  as  stated  below. 

AVERAGE  DAILY  NITROGEN  AND  PHOSPHORUS  BALANCES  WITH  A 
MAN  ON  DIETS  WITH  DIFFERENT  LECITHIN  CONTENTS— Grams 


Nitrogen 

Phosphorus  (P2O5) 

Food 

Urine 

Feces 

Balance 

Food 

Urine 

Feces 

Balance 

17.96 
18.04 

15.717 
16.730 

2.173 

2.187 

+0.07 
-0.88 

2.430 
2.745 

1.840 
2.139 

0.603 
0.576 

-0.013 
+0.031 

3-day  preliminary  period. 
3-day  period;  same  diet  as  above 
plus  15  grm.  lecithin  (total). 

Thus  it  appears  that  the  ingestion  of  15  gm.  commercial  leci- 
thin in  two  days  (as  the  authors  state,  though  the  length  of  the  peri- 
od seems  to  be  three  days)  caused  a  change  of  phosphorus  balance 


292 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


from  -0.013  gm.  P205  to  +0.031  gm.  P205.  At  the  same  time  the 
nitrogen  balance  changed  in  the  opposite  direction,  that  is,  from  a 
positive  to  a  negative  balance. 

Buchmann  (1904)  conducted  balance  experiments  with  two 
human  subjects,  administering  lecithin  as  egg  yolk,  and  in  the  pure 
form.      A  part  of  the  data  are  given  below. 

DAILY    CALCIUM,    MAGNESIUM    AND    PHOSPHORUS    METABOLISM 

WITH  ADULT  HUMAN  BEINGS  AS  AFFECTED  BY 

INGESTION  OF  LECITHIN— Grams 


Subject 

Length 

P2O5 

P2O5, 

CaO 

CaO, 

MgO 

MgO, 

and 

of  period 

intake 

percent 

intake 

percent 

intake 

percent 

Diet 

period 

in  days 

Grams 

retained 

Grams 

retained 

Grams 

retained 

Al 

10 

6.13 

8.56 

5.72 

21.88 

0.43 

5.34 

Without  lecithin. 

A2 

7 

7.06 

21.83 

3.30 

7.26 

0.30 

15.08 

With  egg  yolk. 

A3 

7 

7.04 

6.32 

3.28 

2.94 

0.30 

15.07 

Egg  yolk  and  lecithin. 

Bl 

5 

5.65 

neg. 

5.55 

neg-. 

0.42 

neg-. 

Without  lecithin. 

B2 

5 

7.66 

20.37 

4.27 

31.71 

0.33 

61.25 

With  egg  yolk. 

B3 

5 

6.10 

7.21 

5.19 

9.69 

0.48 

54.36 

Without  lecithin. 

B4 

4 

4.35 

0.69 

2.72 

7.74 

0.44 

43.27 

With  edestin. 

Most  of  this  evidence  tends  to  sustain  the  idea  that  lecithin 
increases  phosphorus  retention,  though  the  result  in  period  A3  is 
negative. 

Voltz  (1905)  showed  in  a  series  of  balance  experiments  on  nitro- 
gen metabolism  with  a  dog  that  in  a  ration  of  meat,  rice,  albumin 
and  lard,  the  replacement  of  one-third  of  the  albumin  nitrogen  by 
the  same  amount  of  lecithin  nitrogen  increased  the  nitrogen  reten- 
tion from  0.020  to  0.140  gm.  per  day. 

Gumpert  (1905)  conducted  balance  experiments  with  sanato- 
gen,  a  preparation  of  casein  and  sodium  glycerophosphate,  with 
results  as  on  the  following  page.  The  subject  in  this  investigation 
was  an  adult  man. 

The  change  in  period  4  from  meat  to  sanatogen,  and  in  period 
5  from  sanatogen  back  to  meat  showed  that  it  was  possible  with  this 
preparation  to  decrease  the  loss  of  phosphorus  and  calcium  existing 
during  the  meat  periods. 

Experiment  2  reports  an  attempt  to  learn  the  effects  of  over- 
feeding with  sanatogen.  By  its  use  it  was  found  possible  to  bring 
about  a  marked  storage  of  both  nitrogen  and  phosphorus,  which 
fact  gains  added  significance  from  the  coincident  loss  in  calcium. 
The  plan  of  the  experiment  is  such,  however,  that  we  are  unable  to 
judge  of  the  participation  of  the  glycerophosphoric  acid  of  the  san- 
atogen in  the  production  of  the  results  noted.  The  food  in  periods 
2,  3  and  4  of  Exp.  2  was  essentially  the  same  as  in  the  meat  periods 
of  Exp.  1  except  for  the  addition  of  60  gm.  of  sanatogen. 


PHOSPHORUS  METABOLISM 


293 


As  bearing  on  matters  of  general  metabolism  it  is  of  interest 
that  phosphorus  may  be  stored  in  a  grown  man,  in  considerable 
quantity,  for  a  number  of  days  at  least,  irrespective  of  the  calcium 
balance. 

DAILY  NITROGEN,  PHOSPHORUS  AND  CALCIUM  BALANCES  ON  DIETS 
CONTAINING  DIFFERENT  PHOSPHORUS  COMPOUNDS 

Grams 


Exp. 

N 

N 

P2O5 

P2O5 

CaO 

CaO 

No. 

Periods 

Diet 

intake 

balance 

intake 

balance 

intake 

balance 

1 

Normal  mixed 

4  days 

+  meat 

11.6 

+0.38 

1.80 

-0.404 

0.265 

-0.339 

2 

Normal  mixed 

5  days 

+  casein 

11.6 

+0.75 

1.885 

-0.018 

0.367 

-0.088 

3 

Normal  mixed 

I 

3  days 
4 

+  meat 
Normal  mixed 

11.6 

+0.45 

1.80 

-0*252 

0.265 

-0.222 

5  days 

+  sanatogren 

11.6 

+0.54 

2.877 

-0.012 

0.303 

-0.182 

5 

Normal  mixed 

1  day 

+  meat 

11.6 

+0.81 

1.80 

—0.425 

0.265 

-0.313  . 

1 

4  days 

Normal  mixed 

12.0 

+1.34 

1.90 

-0.28 

0.165 

-0.265 

2 

Normal  mixed 

II 

3  days 
3 

+  sanatogen 
Normal  mixed 

19.87 

+3.99 

3.91 

+0.552 

0.216 

-0.214 

3  days 

+  sanatogren 

19.87 

+2.04 

3.91 

+0.143 

0.216 

-0.307 

4 

Normal  mixed 

4  days 

+  sanatogren 

19.87 

+3.74 

3.91 

+0.600 

0.216 

-0.244 

Slowtzoff  (1906b)  showed  in  three  balance  experiments  with 
normal  men,  himself  included,  that  the  ingestion  of  lecithin  caused 
nitrogen  and  phosphorus  retention,  and  a  decrease  of  urinary  sul- 
phur, all  of  which  suggest  protein  synthesis. 

According  to  Slowtzoff,  Umikoff  showed  that  the  protein  re- 
serve is  stored  in  the  muscles  and  liver  principally  as  myosin  and 
myostromin  (the  phosphorus-containing  proteins  that  remain  in 
muscle  after  myosin  extraction,  and  which  in  composition  and  sol- 
ubility resemble  the  nucleoalbumins). 

Slowtzoff  found  that  24  hours  after  the  taking  of  food  a  trans- 
formation of  the  myosin  into  myostromin  took  place.  Slowtzoff 
says  that  if  his  conclusions  and  those  of  Umikoff  are  correct  we  may 
consider  the  transformation  of  absorbed  protein  into  the  organized, 
as  an  enriching  of  absorbed  protein  with  phosphoric  acid  and  xan- 
thin  substances.  Then  the  action  of  lecithin  would  be  favorable  for 
this  organization,  and  it  would  be  comprehensible  that  protein  reten- 
tion is  accompanied  by  retention  of  xanthin  and  phosphoric  acid. 

Marfori  (1908a,  1908b)  concluded  that  the  phosphorus  of  syn- 
thetic glycerophosphoric  acid,  if  taken  by  the  mouth,  is  easily  ab- 
sorbed and  assimilated;  that  glycerophosphates  introduced  subcu- 
taneously  are  not  retained,  but  instead  are  quickly  excreted  by  the 
kidneys;  lecithin,  on  the  other  hand,  Marfori  found  to  be  retained 
and  assimilated  whether  introduced  per  os  or  subcutaneously. 


294  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Togami  (1908)  sought  to  determine  with  a  growing  dog  wheth- 
er or  not  ingestion  of  sodium  glycerophosphate,  by  a  healthy  animal, 
would  lead  to  increased  phosphorus  storage.  In  a  preliminary 
period  on  meat  alone  the  dog  was  storing  phosphorus.  Then  during  6 
days  there  was  added  to  the  meat  diet,  each  day,  3.2291  gm.  P205 
in  the  form  of  sodium  glycerophosphate.  This  change  produced 
marked  digestive  disturbance,  with  vomiting  on  the  third  and  fourth 
days.  There  was  a  marked  increase  in  the  urinary  phosphorus, 
and  a  decrease  in  the  phosphorus  retention,  though  the  phosphorus 
balance  remained  positive.  In  the  after  period  of  6  days,  on  meat 
alone,  the  phosphorus  balance  was  negative,  the  daily  urinary  phos- 
phorus excretion  showing  a  gradual  elimination  of  accumulated 
phosphorus  from  the  glycerophosphate. 

There  was,  therefore,  no  evidence  of  permanent  retention  of 
phosphorus  from  the  sodium  glycerophosphate.  Clearly  it  was  not 
retained.  One  should  bear  in  mind,  however,  that  the  poverty  of 
the  basal  ration  in  calcium,  and  the  fact  that  the  glycerophosphate 
was  fed  as  a  sodium  salt  were  conditions  unfavorable  for  phosphorus 
retention. 

J.  and  W.  Cronheim  (1910)  studied  phosphorus  metabolism  in 
infants,  as  affected  by  lecithin,  which  was  used  to  replace  such  part 
of  the  milk  as  to  leave  the  nitrogen  and  phosphorus  of  the  milk  plus 
lecithin  as  nearly  as  possible  the  same  as  of  the  diet  of  milk  alone. 
With  one  child  the  lecithin  increased  the  nitrogen  retention  from 
2.69  percent  of  the  intake  to  4.44  percent  of  the  same,  and  the  phos- 
phorus retention  from  a  negative  balance  to  a  positive  balance  of 
15.69  percent  of  the  intake. 

With  a  second  child  the  nitrogen  retention  was  reduced  from 
5.61  to  4.01  percent  of  the  intake,  and  the  phosphorus  retention 
increased  from  1.69  to  2.34  percent  of  the  intake.  Calcium  reten- 
tion was  decreased  in  both  cases.  These  results  must  be  considered 
inconclusive. 

Yoshimoto  (1910)  studied  the  effects  of  lecithin  ingestion  ori 
protein  metabolism  in  the  dog.  Lecithin  was  added  to  a  basal  ration 
of  horse-flesh,  bacon,  salt  and  water.  In  the  lecithin  periods 
there  was  evidently  a  significant  retention  of  both  nitrogen  and 
phosphorus,  the  effects  continuing  into  the  after-periods. 

A.  Loeb  (1911)  found  in  balance  experiments  with  two  human 
beings  that  the  retention  of  lecithin  phosphorus  was  not  accom- 
panied by  such  a  decrease  of  calcium  and  phosphorus  outgo  as  sug- 
gested deposition  of  the  phosphorus  in  the  bones,  in  fact  the  calcium 
outgo  was  increased.  The  balance  periods  were  3-5  days  in  length 
— insufficient  for  the  purpose. 


PHOSPHORUS  METABOLISM 


295 


Bickel  (1911a)  conducted  balance  experiments  with  a  man  26 
years  old  and  weighing  about  66  kg.,  studying  the  influence  of  leci- 
thin on  metabolism.  The  following  table  we  have  calculated  from 
the  author's  data: 

AVERAGE  DAILY  NITROGEN  AND  PHOSPHORUS   (P205)    BALANCES 
WITH  A  MATURE  MAN  AS  INFLUENCED  BY  LECITHIN— Grams 


Nitrogen 

Phosphorus  (P2O5) 

Periods 

Food 

Urine 

Feces 

Balance 

Food 

Urine 

Feces 

Balance 

Diet 

Fore-period 
3  days 

Main  period 
5  days 

After  period 
5  days 

24.530 

16.387 
19.450 
19.416 

4.200 
3.080 
3.416 

+3.943 
+2.000 
+1.698 

7.880 
9.160 

7.880 

4.200 
4.638 
3.996 

1.633 
2.260 
2.014 

+2.047 
+2.262 
+1.866 

Normal  mixed 

stand  aid  diet. 

Same  plus   "bioci- 
tin." 

Standard  diet  plus 
dry  egg  white. 

Fore-period 
3  days 

Main  period 
5  days 

After  period 
5  days 

16.997 
17.356 

17.444 

4.533 
4.520 

4.404 

+3.000 
+2.654 

+2.682 

7.880 
8.400 

7.880 

3.713 
3.814 

3.642 

1.257 
1.280 

1.474 

+2.910 
+3.306 

+2.764 

Standard  diet  plus 

dry  egg-  white. 
Standard  diet,  dry 

eg-g-  white  and 

lecithin. 
Standard  diet  plus 

dry  egg-  white 

Lecithin  fed  in  the  pure  form,  or  as  "biocitin,"  was  apparently- 
absorbed  and  retained,  at  least  the  feeding  of  these  preparations  in- 
creased the  phosphorus  retention.  The  amounts  of  phosphorus 
ingested  were  about  twice  the  maintenance  requirement,  and  the 
phosphorus  retention  was  remarkably  large.  See  also  Bickel  (1911b). 

Patta  (1912)  reports  that,  in  doses  of  0.50-0.75  gm.  per- day, 
lecithin  manifests  a  sparing  action  toward  the  phosphorus  and  nitro- 
gen balance  of  the  body,  this  effect  being  most  pronounced  when 
there  has  been  a  negative  balance  during  the  period  preceding  the 
injection. 

Satta  and  Gastoldi  (1913)  report  that  if  animals  are  in  nitrogen 
and  phosphorus  equilibrium  they  eliminate  the  phosphorus  of 
lecithin  principally  as  inorganic  phosphate  in  the  urine,  and  that  if 
not  in  equilibrium  phosphorus  from  the  lecithin  is  retained. 

Summary.  The  phosphorus  of  lecithin  and  glycerophosphates 
may  be  absorbed  from  the  alimentary  tract  and  utilized  in  the  tis- 
sues. Lecithin  administered  subcutaneously  may  be  retained,  but 
there  is  some  evidence  that  glycerophosphates,  under  this  condition, 
are  quickly  eliminated  by  the  kidneys. 

Lecithin  added  in  the  form  of  egg  yolk  to  the  milk  diet  of  an 
infant  appears  to  favor  nitrogen  and  phosphorus  retention,  and  gain 
in  weight. 

Egg  yolk  as  compared  with  plasmon— a  milk  albumin  prepara- 
tion— appeared,  from  experiments  with  dogs  and  guinea  pigs,  to 
favor  the  growth  of  the  animal,  and  the  development  of  the  bones. 


296  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

As  with  other  phosphorus  compounds  lecithin  and  the  glycero- 
phosphates are,  in  their  metabolism,  largely  independent  of  nitrogen 
and  calcium,  at  least  during  the  limited  periods  covered  by  most  bal- 
ance experiments. 

We  have  in  these  balance  data  no  evidence  of  the  possession  by 
lecithin  or  glycerophosphates  of  any  unusual  nutritive  or  stimu- 
lative values,  when  added  to  the  normal  food  of  healthy  animals. 
That  they  do  possess  a  higher  degree  of  usefulness  in  states  of  im- 
poverishment, however,  seems  probable.  Naturally,  the  apparent 
results  of  the  administration  of  these  compounds  depend  on  the  nu- 
tritive status  of  the  subject,  and  on  the  other  dietetic  treatments 
used  for  comparison.  In  some  states  of  nutritive  derangement 
they  possess  life-saving  capacity ;  in  some  other  more  favorable  cir- 
cumstances they  may  be  of  no  unusual  value. 

GROWTH  AND  COMPOSITION  OF  ANIMALS  AS  AFFECTED  BY 
COMPOUNDS  OF  GLYCEROPHOSPHORIC  ACID 

Protracted  experiments  with  glycerophosphoric  acid  com- 
pounds in  the  growth  of  animals  have  been  prolific  of  results  of  in- 
terest, and  there  is  considerable  evidence  of  this  nature  which  indi- 
cates at  least  a  high  degree  of  usefulness  of  these  compounds  in  the 
animal  economy,  if  indeed  some  of  them  are  not  essential  to  the 
maintenance  of  life. 

Heffter  (1891)  shows  that  the  lecithin  of  the  liver  is  decreased 
by  starvation  and  by  phosphorus  poisoning. 

Umikoff  (1895)  experimented  with  rats  and  doves  on  rations 
containing  various -compounds  of  phosphorus.  They  throve  only 
on  rations  containing  lecithin. 

Danielewsky  (1895b)  reared  tadpoles  in  lecithin  solutions,  and 
reported  marked  increase  in  growth  in  excess  of  that  made  by  the 
controls  in  water  without  lecithin.  He  ascribes  to  lecithin  a  marked 
stimulating  influence  on  the  processes  of  multiplication  of  cellular 
elements.  Danielewsky's  observations  on  the  effects  of  lecithin  on 
the  growth  of  tadpoles  have  not  been  corroborated. 

Danielewsky  (1896)  conducted  injection  and  feeding  experi- 
ments on  chickens  and  young  dogs  with  lecithin.  In  each  case  the 
animal  receiving  the  lecithin  made  greater  gain  in  weight  than  the 
control.  The  doses,  given  at  intervals  of  3-5  days,  were  about  5-10 
mg.  for  the  chicks,  and  20-50  mg.  for  the  pups.  The  dose  was 
doubled  or  tripled  when  given  by  the  mouth. 

Danielewsky  states  that  lecithin  has  a  stimulating  influence, 
doubtless  in  connection  with  the  increase  of  erythrocytes  and 
haemoglobin  in  the  blood.      Numerical  data  are  given  on  page  297. 


PHOSPHORUS  METABOLISM 


297 


EFFECT  OF  LECITHIN  ON  THE  GROWTH  OF  CHICKS  AND  YOUNG 

DOGS 


Series 

Animals 

First  weight 
Grams 

Final  weight 
Grams 

Treatment 

Time  of 
experiment 

I 

1 

2  (ave.) 

2  (ave.) 

1 

1 

1 

1 

1 

1 

1 

1 

1 

1 

1 

97 

99.5 
249 
264 
495 
560 
746 
679 
495 
484 
459 
487 
670 
720 

785 

746 

1187 

880 

4080 

3630 

4085 

2965 

7450 

11170 

7420 

6820 

6570 

5930 

Lecithin 

Control 
Lecithin 

Control 
Lecithin 

Control 
Lecithin 

Control 
Lecithin 
Lecithin 

Control 

Control 
Lecithin 

Control 

Aug-.    26-Dec.    5 

I 

Aug-.   26-Dec.    5 

II 

Sept.  28-Nov.  13 

II 

Sept.  28-Nov.  13 

Ill 

Aug-.    25-Oct.  30 

Ill 

Aug.    25-Oct.  30 

IV 

June  21-Nov.  28 

IV 

June  21-Nov.  28 

V -. 

June    21-Oct.  18 

V 

June    21-Oct.  18 

V..., 

V 

June    21-Oct.  18 
June    21-Oct.  IS 

VI 

Jan.    25-Apr.  17 
Jan.    25-Apr.  17 

Danielewsky  (1897)  submitted  photographs  showing  lecithin- 
treated  animals  surpassing  the  controls  by  one-half  or  two-thirds  in 
length.  He  also  claims  that  the  lecithin  affects  the  psychic  devel- 
opment of  the  young  dogs  in  a  remarkable  manner. 

Desgrez  and  Zaky  (1900)  injected  subcutaneously  into  guinea 
pigs  lecithin  dissolved  in  sterile  olive  oil.  The  dose  was  40-60  mg.  per 
day  during  8  or  10  days.  The  lecithin  increased  the  urinary  nitro- 
gen, decreased  the  urinary  phosphorus  outgo,  and  increased  the  live 
weight  as  compared  with  controls.  The  apparent  result,  of  course, 
depends  on.  the  treatment  of  the  control. 

Wildiers .  (1900)  reports  negative  results  from  the  feeding  of 
lecithin  to  tadpoles,  chickens  and  dogs.  The  metamorphosis  of  the 
tadpoles  was  delayed ;  no  increased  growth  or  increase  of  red  blood 
corpuscles  was  produced  by  hypodermic  injection  of  lecithin  in  an 
anaemic  dog. 

Carriere  (1901)  reports  benefit  from  the  use  of  lecithin  with 
six  normal  children,  during  a  period  of  six  months.  Data  are  sub- 
mitted on  height  and  weight  before  and  after  treatment ;  and  state- 
ments are  made  as  to  results  on  blood  and  urine,  but  no  data  are  in- 
cluded on  feces  or  food.  According  to  Carriere's  observations  the 
usual  effects  of  lecithin  to  increase  the  urea  and  decrease  the  urin- 
ary phosphorus  were  at  first  apparent,  but  passed  away  by  the  end 
of  the  experiment.  The  red  blood  corpuscles  are  said  also  to  have 
been  increased  by  lecithin  treatment. 

Desgrez  and  Zaky  (1901a,  1902a,  1902b,  1902c)  conducted  a 
series  of  studies  on  the  influence  of  lecithin  on  the  animal  organism, 
especially  on  the  live  weight,  the  development  of  the  skeleton  and 
nervous  tissue,  and  on  the  composition  of  the  urine.  The  subjects 
were  guinea  pigs,  rabbits  and  dogs.  Egg-yolk  lecithin  was  admin- 
istered by  the  mouth,  or  subcutaneously.  A  part  of  the  results  are 
as  follows : 


298  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1.  Three  lots  of  guinea  pigs,  3  in  each  lot,  were  fed  on  bread, 
bran  and  cabbage;  quantities  not  stated.  One  lot  served  as  con- 
trols; a  second  received  lecithin  subcutaneously,  and  a  third  re- 
ceived lecithin  in  pills.  In  43  days  the  controls  gained  480  gm., 
those  treated  subcutaneously  670  gm.  and  those  receiving  lecithin 
per  os  850  gm.  The  two  lots  receiving  lecithin  excreted  more  urin- 
ary nitrogen,  but  less  urinary  phosphorus  than  the  controls. 

2.  Two  guinea  pigs  from  the  same  litter  were  used  as  the  sub- 
jects of  a  similar  experiment ;  one  was  used  as  a  control,  and  the 
other  was  injected  with  lecithin.  In  one  month  the  control  gained 
in  weight  120  gm. ;  the  injected  one  240  gm. 

3.  Of  three  dogs  from  the  same  litter  one  was  used  as  a  con- 
trol; one  was  injected  subcutaneously,  and  the  third  received  the 
same  amount  of  lecithin  in  the  form  of  pills.  In  27  days  the  con- 
trol gained  1480  gm.,  the  injected  dog  2050  gm.  and  the  one  receiv- 
ing lecithin  per  os  2100  gm.  The  dogs  receiving  lecithin  excreted 
more  nitrogen,  but  less  phosphorus  than  the  control. 

4.  During  inanition  it  was  found  with  guinea  pigs  that  those 
injected  with  lecithin  in  olive  oil  lived  longer  than  those  injected 
with  olive  oil  alone. 

5.  After  60  days  treatment  as  under  (1)  it  was  found,  by 
killing  the  guinea  pigs,  that  both  the  absolute  and  relative  weight 
of  the  brains  of  the  treated  guinea  pigs  were  greater  than  the  con- 
trols, while  the  weight  of  the  femora  of  the  treated  animals  ex- 
ceeded the  controls  only  in  the  case  of  those  which  had  received  leci- 
thin by  the  mouth. 

6.  After  7  months'  treatment  as  under  (2) ,  the  control  and  the 
treated  animals  compared  as  follows :  live  weight,  750  gm.  :830  gm. ; 
weight  of  brain,  2.08  gm.:2.24  gm.;  weight  of  left  femur,  1.28 
gm.  :1.60  gm. 

7.  Five  guinea  pigs  were  used  as  controls  while  five  others 
received  lecithin  by  the  mouth,  from  Nov.  1  to  Dec.  24.  All  were 
killed  on  Jan.  6  and  7.  The  controls  and  the  treated  animals  com- 
pared as  follows:  live  weight,  2510  gm.:2840  gm.;  gain  in  body 
weight,  780  gm.:1200  gm. ;  brain,  16.56  gm.:18.14  gm.;  left  femur, 
6.16  gm.  :6.65  gm. ;  phosphorus  per  100  gm.  of  brain,  0.358  gm.  :0.373 
gm. ;  lecithin  per  100  gm.  of  brain,  4.03  gm.  :4.19  gm. ;  mineral  mat- 
ter per  100  gm.  of  femur,  66.80  gm.:69.30  gm.;  P2Og  per  100  gm. 
mineral  matter,  39.68  gm.:41.52  gm. 

8.  Of  2  rabbits  one  was  used  as  a  control,  and  the  other  re- 
ceived daily  0.10  gm.  lecithin  by  the  mouth.  During  40  days  the 
control  gained  200  gm.,  and  the  other  350  gm.  in  live  weight.  The 
control  and  the  lecithin  rabbit  compared  as  follows:  live  weight, 


PHOSPHORUS  METABOLISM  299 

2020  gm.:2200  gm.;  weight  of  brain,  8.18  gm.:9.28  gm.;  weight  of 
femur,  8.26  gm.  :8.59  gm. ;  total  phosphorus  per  100  gm.  of  brain, 
0.341  gm. :0.367  gm. ;  mineral  matter  per  100  gm.  femur,  61.74  gm.: 
62.20  gm.;  P205  per  100  gm.  mineral  matter  of  femur,  38.01  gm. : 
39.91  gm. 

9.  Of  two  rabbits  from  the  same  litter,  one  was  used  as  a  con- 
trol, and  the  other  received  0.10  gm.  lecithin  daily  for  two  and  a 
half  months.  The  control  and  the  lecithin  rabbit  compared  as  fol- 
lows: live  weight,  2450  gm.  :2170  gm. ;  gain  in  weight,  760  gm.  :890 
gm. ;  brain,  8.92  gm.  :9.11  gm. ;  femur,  10.14  gm.  :8.28  gm. ;  phospho- 
rus per  100  gm.  brain,  0.356  gm.  :0.347  gm. ;  mineral  matter  per 
100  gm.  femur,  65.10  gm.  :66.94  gm. ;  P205  per  100  gm.  mineral  mat- 
ter of  femur,  37.17  gm.  :39.71  gm. 

10.  Of  two  dogs  of  the  same  age,  one  was  used  as  a  control, 
and  the  other  received  0.10  gm.  lecithin  per  day  from  Oct.  1  to  Dec. 
6.  The  control  and  the  lecithin  dog  compared  as  follows:  live 
weight,  2550  gm.  :3780  gm. ;  gain  in  weight,  300  gm.  :1380  gm. ; 
weight  of  brain,  46.42  gm.  ;49.90  gm. ;  weight  of  femur,  10.27 
gm.  :11.00  gm. ;  phosphorus  per  100  gm.  of  brain,  0.365  gm.  :0.397 
gm. ;  lecithin  per  100  gm.  of  brain,  3.73  gm.  :4.06  gm. ;  mineral  mat- 
ter per  100  gm.  femur,  61.03  gm.  :62.81  gm. ;  P205  per  100  gm.  min- 
eral matter  in  femur,  38.9  gm. : 37.86  gm. 

11.  Of  two  dogs  of  the  same  litter,  one  was  used  as  a  control, 
while  the  other  received  daily  for  two  months  0.10  gm.  lecithin. 
Three  months  later  the  dogs  were  killed.  The  control  and  the 
lecithin  dog  compared  as  follows :  live  weight,  13300  gm. :  13340  gm. ; 
weight  of  brain,  69  gm.  :76  gm. ;  weight  of  femur,  59  gm.  :68  gm. ; 
phosphorus  per  100  gm.  brain,  0.357  gm.  :0.340  gm. ;  lecithin  per  100 
gm.  brain,  3.82  gm. :  3.97  gm. ;  mineral  matter  per  100  gm.  femur, 
62.40  gm.  :65.14  gm. ;  P205  per  100  gm.  mineral  matter  in  femur, 
39.07  gm.  :39.82  gm. 

12.  In  analyzing  the  mode  of  action  of  lecithin,  Desgrez  and 
Zaky  found  with  guinea  pigs  that  sodium  glycerophosphate  in- 
creased the  urinary  nitrogen,  but  did  not  alter  the  phosphorus  out- 
go. Choline,  however,  subcutaneously  injected  in  quantities  of  1 
c.c.  of  a  2-percent  solution  during  8  months,  decreased  the  urinary 
phosphorus,  and  increased  the  live  weight.  Betain  also  decreased 
the  urinary  phosphorus,  but  caused  loss  in  live  weight. 

A  part  of  the  author's  conclusions  are  as  follows: 
Lecithin  of  egg  exercises  on  the  nutritive  exchanges  a  favorable 
influence  which  is  manifested  by  an  increase  in  the  urea,  the  total 
nitrogen,  and  the  nitrogen  coefficient.  There  is  at  the  same  time 
a  constant  decrease  in  the  amount  of  phosphoric  acid  eliminated 
through  the  urine.  The  appetite  and  weight  of  the  animals  in- 
crease. 


300 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


Lecithin  administered  to  guinea  pigs  in  a  state  of  inanition  pro- 
longs life  some  days.  The  loss  of  weight  at  the  time  of  death  is 
practically  equal  in  both  control  and  treated  animals;  the  propor- 
tion of  material  utilized  is  the  same ;  the  process  of  nutrition  being 
better  in  the  treated  animals,  they  survive  longer,  because  of  a  more 
perfect  elaboration  of  their  reserves  and  of  the  protein  of  their  tis- 
sues. 

The  increase  in  weight  of  the  treated  animals  does  not  mean 
an  increase  in  fatty  tissue.  It  takes  place  proportionately  in  the 
skeleton  and  the  nervous  system.  In  the  skeleton  it  results  in  an 
increasing  of  the  mineral  material,  particularly  of  phosphoric  acid ; 
in  the  nervous  system,  an  increase  in  total  phosphorus  and  in  leci- 
thin. 

Springer  (1902)  reports  lecithin  injection  and  feeding  experi- 
ments with  guinea  pigs,  rabbits  and  dogs.  The  results  were 
inconclusive  as  to  effects  on  weight  of  brain  per  unit  of  body  weight, 
and  also  on  weight  and  length  of  bones.  The  total  phosphorus  and 
the  lecithin  of  the  brain,  and  the  mineral  matter  of  the  bone  were 
in  each  case  greater  where  lecithin  had  been  administered  than  in 
the  controls.      A  part  of  the  figures  are  as  follows : 

EFFECTS  OF  LECITHIN  ON  THE  COMPOSITION  OF  THE  BRAIN  AND 

BONES  OF  ANIMALS 


Experiment 
and  subject 

Method  of  adminis- 
tration of  lecithin 

Percent 

phosphorus 

(P2O5)  in  brain 

Percent 
lecithin 
in  brain 

Percent 

mineral  matter 

in  femur 

Percent  P2O5 

in  mineral 
matter  of  femur 

II 
Guinea  pigs... . 

Controls  (5) 
0.05  gm.  in  food  daily 

0.358 
0.373 

4.03 
4.19 

66.80 
69.30 

39.68 
41.52 

Ill 

Control  (1) 
0.100  gm.  daily  in  food 

0.341 
0.367 

61.74 
66.20 

38.01 
39.91 

IV 

Control  (1) 
0.100  gm.  daily  in  food 

0.365 
0.397 

3.73 
4.06 

61.03 
62.81 

38.90 
37.86 

Hatai  (1903)  conducted  feeding  and  injection  experiments  with 
lecithin  on  white  rats.  The  treated  animals  in  each  of  5  cases 
gained  more  in  weight  than  the  controls,  though  the  amounts  of 
food  consumed  were  not  stated.  The  relative  weight  of  the  cen- 
tral nervous  system  of  the  lecithin  rats  was  normal,  as  was  also  its 
moisture  content  and  the  relative  development  of  sheath  and  axis 
cylinder. 

Lusena  (1903)  determined  that  the  lecithin  content  of  the  liver, 
kidneys  and  myocardium  under  acute  poisoning  by  arsenic  and  by 
phosphorus  did  not  vary  much  from  the  normal,  and  concluded 
(a)  that  in  experimental  fatty  degeneration  the  larger  part  of  the 
fat  in  the  degenerated  organs  is  infiltrated  fat,  (b)  that  it  is  not  dis- 


PHOSPHORUS  METABOLISM  301 

proved  that  a  part  of  the  fat  is  of  endocellular  origin,  from  the 
transformation  of  protein,  and  (c)  that  though  the  protein  may- 
change  to  fat,  the  lecithin  is  not  the  transformation  substance,  for 
the  lecithin  is  present  within  normal  variations  both  in  cloudy  swell- 
ing and  advanced  fatty  degeneration.  See  also  Billon  and  Stassano 
(1903a,  1903b). 

Dorn  (1904)  conducted  feeding  experiments  with  rabbits  show- 
ing the  nutritive  value  of  lecithin  and  protylin.  Protylin  is  recom- 
mended for  use  in  scrofula,  anaemia  and  rachitis. 

Desgrez  and  Zaky  (1904a,  1904b,  1904c,  1905)  compared  leci- 
thin, protylin,  nuclein  (from  yeast),  and  nucleic  acid  (also  from 
yeast) ,  as  to  their  influence  on  the  development  and  composition  of 
animals.  The  subjects  were  dogs  and  guinea  pigs,  and  the  program 
included  feeding  experiments,  with  some  metabolism  data,  and  par- 
tial body  analyses. 

All  of  these  organic  phosphorus  compounds  were  found  to  be  of 
value  in  causing  increase  in  weight  greater  than  that  of  the  controls. 
The  brain  increased  in  weight,  as  did  the  bones ;  and  the  bones  in- 
creased in  ash  under  the  influence  of  these  compounds.  The  auth- 
ors considered  lecithin  and  protylin  to  be  more  valuable,  and  nucleic 
acid  less  .valuable  than  the  other  compounds  in  causing  gain  in  live 
weight.  Since  there  were  no  feces  phosphorus  figures  we  have  no 
basis  for  a  critical  judgment  of  their  conclusions. 

Pignatti  (1906)  studied  the  influence  of  oral  or  subcutaneous 
introduction  of  various  organic  phosphorus  compounds  on  the  fer- 
ratin  in  the  liver.  Casein,  sodium  glycerophosphate,  and  lecithin 
increased  the  ferratin  of  the  liver.  It  was  further  concluded  that 
by  injection  of  the  glycerophosphate  the  phosphorus  content  of  fer- 
ratin is  not  increased,  but  that  casein  injection  does  give  a  ferratin 
possessing  a  high  phosphorus  content. 

Morgen,  Beger  and  Fingerling  (1906)  report  results  of  sheep 
and  goat  feeding  experiments,  with  lecithin,  as  affecting  milk  pro- 
duction, which  seem  to  show  that  in  doses  of  1-2  gm.  this  compound 
increases  the  secretion  of  milk  and  milk  solids,  fat  production,  how- 
ever, being  increased  only  when  the  basal  ration  was  low  in  fat. 

Goldfarb  (1907)  studied  the  effects  of  lecithin  on  growth  with 
tadpoles  and  kittens.  The  tadpoles  were  kept  in  lecithin  solutions  of 
various  strength,  from  one  one-hundred-fiftieth  to  two  percent.  The 
kittens  received  lecithin  subcutaneously,  or  in  the  food.  The  tad- 
poles so  treated  showed  no  greater  increase  in  weight  than  the  con- 
trols. The  kittens  which  received  the  lecithin  gained,  on  an  aver- 
age, about  7  percent  more  than  the  controls. 


302  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

In  a  later  series  of  experiments  (1910)  similar  negative  results 
were  obtained  with  tadpoles  and  sea-urchin  eggs,  also  in  lecithin  sol- 
ution ;  with  an  experiment  involving  52  kittens,  those  receiving  leci- 
thin did  not,  on  the  whole,  exceed  in  growth  those  not  receiving 
lecithin.  An  experiment  on  a  litter  of  5  guinea  pigs  also  gave  neg- 
ative results. 

Franchini  (1907, 1908a)  investigated  the  question  of  method  of 
utilization  of  lecithin  by  the  animal  organism,  in  experiments  on  rab- 
bits. Two  lots  of  seven  rabbits  each  were  used.  During  three  days 
each  rabbit  of  one  lot  received  4.799  gm.  of  pure  lecithin,  but  no 
other  food.  The  other  lot  received  neither  lecithin  nor  food.  Both 
lots  received  water  to  drink.  Both  lots  were  killed  four  days  after 
the  beginning  of  the  treatment,  twenty-four  hours  after  the  last 
feeding  of  lecithin. 

The  lecithin  feeding  increased  the  lecithin  content  of  the  liver 
and  muscle,  but  not  of  the  brain.  Franchini  accepts  SlowtzofFs 
conclusion  that  lecithin,  in  part  at  least,  is  absorbed  as  such,  unsplit. 

Forbes  (1909)  compared  lecithin  with  other  organic  and  inor- 
ganic phosphorus  compounds  in  the  growth  of  swine.  Results  on 
more  individuals  will  be  necessary  to  settle  many  of  the  points  of 
interest,  but  the  observed  effects  of  lecithin  on  the  composition  of 
the  muscles  is  considered  characteristic.  The  moisture  content  of 
the  fat-free  muscle  was  higher  than  in  any  other  lot,  as  also  was  the 
total  phosphorus,  either  as  related  to  the  whole  meat,  the  protein  or 
the  ash.  See  also  Forbes  (1909) ;  Common  Foods  in  Rel.  to  P.  Met. 
and  Forbes  and  associates  (1914) ;  Nutr.  Val.  Org.  and  Inorg.  P. 

W.  Cronheim  (1912)  conducted  metabolism  experiments,  with 
two  mature  men  on  rations  varying  in  lecithin  content,  from  which 
he  concluded  that  for  grown  people  as  well  as  children  lecithin  is  an 
important  constituent  of  the  diet,  and  that  it  has  the  same  value 
for  adults  as  has  often  been  observed  for  children,  in  favoring  the 
replacement  of  lost  nitrogenous  material. 

Wesselkin  (1913)  made  a  microchemical  study,  with  rabbits  as 
subjects,  of  the  fatty  bodies  deposited  in  the  organs  as  a  result  of 
the  feeding  of  lecithin  and  of  egg  yolk.  Both  forms  of  nutriment 
caused  a  deposit  of  lipoid  substances  other  than  true  fats.  Among 
these  substances  were  phosphatids  and  apparently  lecithin.  The 
phosphatid  formed  as  a  result  of  the  yolk  feeding  was  much  in  ex- 
cess of  the  amount  stored  as  a  result  of  the  feeding  of  the  same 
amount  of  lecithin  in  the  pure  form.  The  lipoids  resulting  from 
feeding  egg  yolk  were  mainly  cholesterin  esters. 


PHOSPHORUS  METABOLISM  303 

Salkowski  (1913b)  reports  an  increase  of  the  phosphorus  of  the 
brain  through  -the  administration  of  cephalin  in  pill  form  to  fasting 
rabbits  during  four-day  periods.  It  is  Salkowski's  idea  that  ceph- 
alin is  stored  in  the  brain.  The  extent  of  the  work  was  insufficient 
to  establish  so  important  a  point. 

Summary.  In  all  such  experiments  as  the  above  the  results 
depend  on  the  comparison  of  the  experimental  animals  with  the  con- 
trols, that  is,  of  the  animals  receiving  the  nutrients  of  interest,  with 
others  not  receiving  them.  It  is,  therefore,  obvious  that  the  status 
of  the  controls  has  as  much  to  do  with  apparent  results  as  does  the 
behavior  of  the  animals  receiving  the  nutrients  in  question. 

It  is  our  own  belief,  therefore,  that  some  at  least  of  the  differ- 
ences in  the  conclusions  of  the  different  investigators  are  due  to 
differences  in  the  conditions  attending  the  experiments,  especially 
in  the  state  of  nutrition  of  the  animals,  and  their  susceptibility 
to  the  effects  of  the  experimental  treatment,  as  determined  by  pre- 
vious conditions  of  life.  It  would  therefore  be  desirable  that  we 
know  much  more  than  has  been  recorded  in  regard  to  some  of  the 
conditions  which  affect  results,  but  which  are  not  commonly  recog- 
nized as  of  importance. 

From  the  investigations  above  mentioned  one  must  conclude 
that  whatever  the  function  of  lecithin  and  related  compounds  in  the 
normal  life  of  animals  there  are  conditions  not  definitely  distinct 
from  the  normal  in  which  they  do  possess  certain  important  specific 
effects. 

In  comparison  with  more  or  less  carefully  chosen  controls,  ani- 
mals receiving  lecithin,  especially  if  the  basal  ration  be  low  in  phos- 
phorus, and  the  animals  in  a  state  of  depleted  reserves,  show  in- 
creased appetite  and  gain  in  live  weight;  phosphorus  retention  is 
favored ;  there  has  been  observed  an  increase  of  ash  and  of  phospho- 
rus in  the  skeleton,  and  of  phosphorus  in  the  muscles,  and  of  phos- 
phorus and  lecithin  in  the  brain ;  fasting  animals  live  longer  if  they 
receive  lecithin,  and  their  livers  and  muscles  are  increased  in  lecithin 
content  by  ingestion  of  lecithin.     See  also  Lecithin  Therapy. 

METABOLISM  OF  PHOSPHOCARNIC  ACID 

Siegfried  (1894,  1895,  1896)  argues  that  since  phosphocarnic 
acid  contains  both  phosphorus  and  iron,  and  readily  forms  easily  sol- 
uble compounds  with  lime  and  magnesia,  and  since  these  compounds 
are  soluble  in  either  neutral,  weakly  acid  or  weakly  alkaline  solu- 
tions it  is  fitted  to  serve  as  a  carrier  of  phosphorus,  iron,  lime  and 


304 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


magnesia  throughout  the  body  fluids.  Its  presence  in  considerable 
quantity  in  milk  is  in  harmony  with  this  view.  Taken  in  meat  or 
its  extractives  it  can  render  available  the  lime  either  of  other  foods 
or  of  drinking  water. 

Siegfried  performed  experiments  on  dogs  in  a  study  of  the  re- 
lation of  phosphocarnic  acid  to  muscular  work.  A  nerve  in  one 
hind  leg  was  cut ;  then  the  corresponding  muscle  of  the  other  hind 
leg  was  stimulated  to  activity  for  one  hour  by  an  electric  current. 
The  dog  was  then  killed,  and  phosphocarnic  acid  determined  in  the 
fresh  and  in  the  tired  muscle.  The  tired  muscle  always  showed  less 
phosphocarnic  acid  than  the  rested  one,  at  one  time  reaching  a  ratio 
of  1:3.  Siegfried  suggests  that  the  phosphocarnic  acid  of  muscle  is 
an  energy-producing  nutrient  rather  than  a  metabolic  waste  pro- 
duct. 

In  the  latest  experiments  described  the  ischiatic  and  crural 
nerves  were  severed,  and  then  stimulated  by  an  induction  current, 
causing  tetanus  alternately  in  the  flexor  and  extensor  muscles.  Af- 
ter 1  hour  and  50  minutes  the  dog  was  killed  and  the  flexors  and  ex- 
tensors of  both  thighs  were  removed  and  examined.  In  a  second  ex- 
periment the  same  nerves  were  severed,  and,  after  the  healing  of 
the  wound  the  dog  was  exercised  rapidly  for  1  hour  and  15  minutes; 
then  killed,  and  the  muscles  examined.  A  third  experiment,  similar 
to  the. second  was  conducted,  the  time  of  exercise  being  1  hour  and 
30  minutes.      Below  are  the  numerical  results. 


PHOSPHOCARNIC  ACID  IN  TIRED  AND  RESTED  MUSCLES 

— Grams 

Weight  of 
muscle 

Absolute  weight  of 
phosphocarnic  acid 
(reckoned  as  carnic) 

Phosphocarnic  acid 
in  1000  grn,  muscle 

Phosphocarnic  acid 

used  for  the  muscular 

work 

Rested 

Tired 

Rested 

Tired 

Rested 

Tired 

Per  1000  gm. 
muscle 

Per  100  parts 

phospho- 
carnic acid 

I., 

ii 

in 

645 
845 
955 

640 
1010 
1210 

1.6193 
1.1116 
0.5463 

0.5996 
0.7344 
0.4697 

2.40 
1.31 

0.57 

0.93 
0.73 
0.39 

1.47 
0.58 
0.18 

61.3 
44.3 
31.6 

The  work  of  Macleod  does  not  sustain  Siegfried's  theory.  See 
Macleod  (1899),  Effects  of  Exercise  on  P.  Met. 

Martin  Miiller  (1897)  found  very  much  lov/er  values  for  phos- 
phocarnic acid  in  the  muscle  in  new-born  infants  than  in  adults. 

Tarozzi's  observations  (1899a,  1899b)  do  not  indicate  any 
change  in  the  proportion  of  phosphocarnic  acid  in  striated  muscle 
during  fast.      See  table  on  next  page. 

Bonanni  (1902)  found  that  the  phosphocarnic  acid  of  the  mus- 
cles of  rabbits  in  acute  veratrum  poisoning  was  reduced  from  the 
normal  1.819  percent  (mean  of  3  determinations)  to  1.426  percent 
(mean  of  4  determinations). 


PHOSPHORUS  METABOLISM 


305 


Cavazzani  (1904a)  studied  the  phosphocarnic  acid  content  of 
the  brains  Of  dogs  after  different  degrees  of  nervous  excitement. 
Those  killed  immediately  after  morphine  sleep  showed  0.217 — 0.285 
percent  ferrinucleon,  with  6.61 — 7.04  percent  nitrogen,  and  those 
killed  after  absynth  excitement  showed  0.543 — 0.690  percent  ferri- 
nucleon, with  3.24 — 5.74  percent  nitrogen. 

PHOSPHOCARNIC  ACID  OF  MUSCLES  FROM  NORMAL  AND  FASTED 

DOGS  (Tarozzi,  1899) 


Duration 
of  fast 

Days 

Loss  of 
weight 

Percent 

Mass  of 
muscle 

Grams 

Carnic  acid 

Condition  of  dog 

In  toto 

Parts  per 
1000 

'6 
28 
50 

io 

28 
45 

400 
450 
320 
435 
595 
442 

0.618 

0.6282 

0.5728 

0.607 

0.787 

0.804 

1.545 

1.396 

1.790 

Fasted 

1.580 

1.324 

1.818 

Panella  (1906a)  found  that  the  quantity  of  phosphocarnic  acid 
in  the  brain  (dog)  gradually  fell  off  during  fast,  the  decrease  being 
in  direct  proportion  to  the  duration  of  the  fast.  The  same  was  true 
independent  of  the  water  content,  which  increased  during  the  fast 
when  water  but  no  food  was  taken. 

METABOLISM  OF  PHYTIN 

FEEDING  EXPERIMENTS  WITH  MEN  AND  ANIMALS 

As  one  of  the  most  important  organic  phosphorus  compounds 
in  foods  of  vegetable  origin,  and  as  that  one  which  contains  much 
the  highest  percentage  of  phosphorus,  especial  attention  attaches 
to  the  metabolism  of  phytin.  A  considerable  amount  of  careful 
work  has  been  done  on  phytin  within  the  past  few  years,  and  its 
status  as  a  nutrient  is  fairly  well  established. 

Scofone  (1904)  found  that  phytin  phosphorus  is  mostly  ab- 
sorbed, and  is  excreted  largely  by  the  kidneys,  as  inorganic  phos- 
phate. 

Giacosa  (1904)  fed  phytin  to  a  dog.  The  phosphorus  of  both 
urine  and  feces  was  increased,  the  inorganic  phosphorus  of  the 
feces  being  increased  more  than  the  organic.  In  administering 
phytin  to  human  beings  (1905),  in  10  gm.  doses,  no  evidence  was 
noted  of  urinary  elimination  of  either  phytin  or  inosite.  The  re- 
lation of  phytin  to  glycogen  was  studied  by  introducing  phytin  into 
the  stomach  of  starving  dogs  and  rabbits.  The  animals  which  re- 
ceived the  phytin  lost  the  most  in  weight,  and  their  livers  con- 
tained, as  a  rule,  less  glycogen.  Giacosa  (1907)  found  the  mortal 
dose  much  less  when  injected  than  when  taken  by  the  mouth. 


806 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


Maestro  (1905b)  presented  balance  data  with  rabbits  showing 
that  phytin  phosphorus  is  absorbed,  and  may  be  retained. 

Gilbert  and  Posternak  (1905)  reported  a  balance  experiment  on 
a  man,  in  an  overfed  condition,  showing  that  the  addition  of  phytin 
to  the  food  increased  the  phosphorus  but  decreased  the  nitrogen  re- 
tention.     Data  on  this  test  are  given  below. 


AVERAGE  DAILY  PHOSPHORUS  METABOLISM  WITH  A  MAN  IN  AN 

OVERFED  CONDITION  AS  AFFECTED  BY  PHYTIN 

Periods  of  Five  Days — Grams 


Nitrogen 

P2O5 

Remarks 

Food 

Urine 

Feces 

Balance 

Food 

Urine 

Feces 

Balance 

22.73 
22.73 

19.01 
20.93 

1.69 
1.51 

+2.03 
+0.29 

3.68 

4.28 

2.721 
2.960 

0.924 
1.262 

+0.039 
+0.062 

Diet     same  as  above    plus  0.6 
gm.  P2O5  daily  as  phytin.  .. 

In  another  experiment  these  authors  compared  phytin  phospho- 
rus with  other  compounds  (see  Nutr.  Val.  Org.  and  Inorg.  P.) ;  but 
the  amounts  fed  were  not  nearly  enough  the  same  to  warrant  con- 
clusions other  than  that  the  phytin  phosphorus  was  absorbed  and 
retained,  but,  as  before,  without  beneficial  effect  on  the  nitrogen 
balance,  which  remained  negative. 

Jordan,  Hart  and  Patten  (1906)  conducted  phosphorus  metabo- 
lism experiments  on  two  milch  cows,  one  animal  being  used  for  Ex- 
periment I  and  the  other  for  Experiments  II  and  III.  The  phos- 
phorus contents  of  the  rations  were  varied  by  choice  of  food  and  by 
method  of  preparation,  as  in  the  washing  of  bran  to  remove  soluble 
phosphorus  compounds.  The  cows  were  kept  in  a  warmed  room  and 
the  excreta  were  caught  by  attendants.  The  cows  were  milked  at 
7  a.  m.,  noon,  five  P.  M.,  and  midnight.  Inorganic  phosphorus  was 
determined  by  the  method  of  Hart  and  Andrews.  Nuclein  and  nu- 
cleo-proteid  phosphorus  were  estimated  as  that  portion  of  the  total 
phosphorus  which  was  insoluble  during  a  15-minute  extraction  with 
0.2  percent  hydrochloric  acid.  Soluble  organic  phosphorus  is  con- 
sidered to  be  the  total  dissolved  by  0.2  percent  hydrochloric  acid 
minus  the  inorganic  phosphorus  as  determined  by  the  Hart  and  An- 
drews method. 

Among  the  authors'  conclusions  are  the  following: 
The  amount  of  outgoing  phosphorus  rose  and  fell  with  the 
quantity  supplied  in  the  food,  though  within  narrower  limits.  When 
the  phosphorus  supply  was  abundant,  there  was  a  storage  of  this 


PHOSPHORUS  METABOLISM  307 

element  in  the  bodies  of  the  animals,  but  during  prolonged  periods, 
in  which  the  phosphorus  supply  was  deficient,  there  was  withdrawn 
from  the  body  store  about  10  gm.  daily. 

Through  katabolic  changes  the  phosphorus  of  the  phytin  and 
that  of  the  unused  digested  nucleo-bodies  was  reduced  to  inorganic 
combinations,  and  was  excreted,  chiefly  in  the  feces,  though  to  a 
small  extent  in  the  urine.  The  inorganic  phosphates  of  the  milk 
were  from  three  to  five  times  greater  in  quantity  than  the  total 
amount  of  such  compounds  in  the  food. 

The  rise  and  fall  in  the  amounts  of  outgoing  phosphorus  com- 
pounds occurred  almost  wholly  with  the  inorganic  salts  found  in  the 
egesta.  The  organic  phosphorus  bodies  of  the  egesta  were  but  lit- 
tle affected,  if  at  all,  by  the  proportions  of  phosphorus  compounds  in 
the  food.  Variations  in  the  phosphorus  supply  appeared  not  to 
modify  the  appropriation  of  this  element  by  the  milk. 

No  relation  whatever  appears  to  exist  between  the  nitrogen  ex- 
cretion and  the  phosphorus  excretion. 

It  was  shown,  without  question,  that  the  physiological  effect  of 
the  two  rations,  due  to  the  withdrawal  from  the  bran  of  such  com- 
pounds as  were  soluble  in  slightly  acidulated  water,  differed  to  a 
marked  degree.  With  the  washed-bran  ration  as  compared  with  the 
one  containing  the  unwashed  bran,  the  following  differences  were 
observed : 

a.  Drier  and  much  firmer  feces  with  the  washed-bran  ration. 

b.  A  greatly  reduced  flow  of  urine  following  a  change  from 
the  unwashed-bran  to  the  washed-bran  ration,  the  reverse  taking 
place  when  a  reverse  change  was  made. 

c.  An  increase  in  the  flow  of  milk  consequent  upon  the  with- 
drawal from  the  ration  of  the  phytin  and  other  water-soluble  con- 
stituents of  bran. 

d.  A  reduction,  sometimes  large,  in  the  percentage  of  fat  in 
the  milk  consequent  upon  the  withdrawal  from  the  ration  of  phytin 
and  other  water-soluble  constituents  of  bran. 

e.  A  decreased  production  of  butter  fat  during  the  period  in 
which  the  washed-bran  ration  was  fed,  notwithstanding  a  some- 
what increased  flow  of  milk. 

The  following  tables  set  forth  some  of  the  main  points  in  this 
work. 


308 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


DAILY  PHOSPHORUS  METABOLISM  WITH  MILCH  COWS 

EXPERIMENT  I 


Total 
P 
fed 


Bails'-  income  and  outgo 


Fed     Outgo     Feces      Milk       Urine 


Rations 


Total  Phosphorus 


Mar.  10-Mar.  16 

12.8 

22.7 

8.40 

14.2 

0.10 

No.  1 
lbs. 

Oat  straw  10  lbs.,  washed  branlO 
,  rice  6  lbs.,  wheat  gluten  1.5  lbs. 

Apr.  12- Apr.  18 

78.7 

70.6 

55.70 

11.0 

3.90 

No.  2 
lbs. 
lib 

No.  1 

Oat  straw  10  lbs.,  whole  bran  10 
,  hominy  feed  5  lbs.,  wheat  gluten 

Apr.  28-May  1 

16.0 

23.8 

13.29 

10.1 

0.37 

Low  phytin  and  nuclein  ration. 

May  9-May  15 

m.H 

70.4 

56.70 

8.8 

4.88 

No.  2 

High  ;;      ;;     ;; 

May  22-May  26 

21.4 

22.7 

9.59 

13.0 

0.11 

No.  1 

Low 

June  10- June  16 

15.1 

20.5 

9.40 

11.0 

0.12 

No.  1 

Low 

Nucleo-phosphorus 


Mar.  10-Mar.  16 
Apr.  12-Apr.  18 
Apr.  28-May  1 
May  9-May  15 
May  22-May  26 
June  10- June  16 


12.8 

7.6 

6.6 

3.9 

2.6 

0.00 

78.7 

24.4 

9.5 

7.6 

1.9 

0.00 

16.0 

9.0 

8.7 

6.9 

1.8 

0.00 

83.3 

28.6 

7.5 

5.8 

1.7 

0.00 

21.4 

14.2 

7.7 

5.2 

2.4 

0.00 

15.1 

7.5 

7.8 

5.2 

2.6 

0.00 

No.  1 
No.  2 
No.  1 
No.  2 
No.  1 
No.  1 


Low 

High 

Low 

Sigh 

Low 

Low 


Soluble 

organic  phosphorus 

Mar.  10-Mar.  16 

12.8 

2.5 

2.0 

1.80 

0.37 

0.00 

No.  1     Low 

Apr.  12-Apr.  18 

78.7 

51.1 

4.1 

3.90 

0.16 

0.00 

No.  2     High    " 

Apr.  28-May  1 

16.0 

4.3 

2.0 

1.40 

0.56 

0.00 

No.  1     Low 

May  9-May  15 

83.3 

52.1 

6.2 

5.30 

0.95 

0.00 

No.  2     High 

May  22-May  26 

21.4 

3.7 

2.2 

1.70 

0.52 

0.00 

No.  1     Low 

June  10-Junel6 

15.1 

4.1 

1.6 

0.98 

0.64 

0.00 

No.  1     Low 

Inorganic  phosphorus 


Mar.  10-Mar.  16 
Apr.  12-Apr.  18 
Apr.  28-May  1 
May  9-May  15 
May  22-May  26 
June  10-June  16 


12.8 

2.63 

14.2 

2.8 

11.3 

0.10 

78.7 

2.60 

56.6 

44.1 

8.6 

3.90 

16.0 

2.60 

13.1 

4.9 

7.9 

0.37 

83.3 

2.60 

57.2 

45.6 

6.7 

4.90 

21.4 

3.50 

12.7 

2.6 

10.0 

0.11 

15.1 

3.40 

11.9 

3.2 

8.6 

0.12 

No.  1 
No.  2 
No.  1 
No.  2 
No.  1 
No.  1 


Low 

High 

Low 

High 

Low 

Low 


Weight  of  cow  1100  lbs.,  3  months  advanced  in  lactation  period, 
corrected  with  approval  of  one  of  the  authors. 


Dates  of  last  period 


PHOSPHORUS  METABOLISM 


309 


DAILY  PHOSPHORUS  METABOLISM  WITH  MILCH  COWS 

EXPERIMENT  II 


Dates 


Total 
P 
fed 


Daily  income  and  outgo 


Outgo     Feces      Milk      Urine 


Rations 


Total  phosphorus 

Dec.  27-Jan.  2 

37 

42.0 

24.4 

17.5 

0.12 

Jan.  13-Jan.  19 

18 

30.2 

11.6 

18.5 

0.09 

Jan.  27-Feb.  2 
Feb.  10-Feb.  16 

37 
20 

39.8 
29.2 

22.1 

10.9 

17.6 

18.2 

0.07 
0.07 

No.  1   Oat  straw  10  lbs.,  washed  bran  10 

lbs.,  corn  germ  meal  6  lbs.,  rice  meal  3 

lbs. 

No-  2    Oat  straw  10  lbs.,  washed  bran  10 

lbs.,  wheat  gluten  2  lbs.,  rice  meal  7  lbs. 

No.  1    High  nucleo-phosphorus. 

No.2    Low        "  'f 


Nucleo-phosphorus 


Dec.  27-Jan.  2 

37 

21.5 

11.4 

8.0 

3.4 

0.00 

No.  1 

High      " 

Jan.  13-Jan.  19 

18 

9.5 

9.7 

6.1 

3.6 

0.00 

No.  2 

Low 

Jan.  27-Feb.  2 

37 

21.2 

11.4 

7.9 

3.5 

0.00 

No.  1 

High      " 

Feb.  10-Feb.  16 

20 

11.4 

10.5 

6.8 

3.7 

0.00 

No.  2 

Low 

Soluble 

organic  phosphorus 

Dec.  27-Jan.  2 
Jan.  13-Jan.  19 
Jan.  27  Feb.  2 
Feb.  10-Feb.  16 

37 
18 
37 
20 

11.1 
4.1 

11.2 
4.8 

3.3 
3.2 
3.2 
2.9 

1.4 
1.4 
1.5 
0.7 

1.9 
1.8 
1.7 
2.2 

0.00 
0.00 
0.00 
0.00 

No.  1    High      ' ' 
No.  2    Low        ' ' 
No.  1    High      ' ' 
No.  2    Low 

Inorganic  phosphorus 


Dec.  27-Jan.  2 
Jan.  13-Jan.  19 
Jan.  27-Feb.  2 
Feb.  10-Feb.  16 

37 
18 
37 
20 

4.7 
4.7 

4.7 
4.7 

27.3 
17.3 
25.3 
15.8 

15.0 
4.1 

12.7 
3.4 

12.1 

13.1 
12.5 
12.3 

0.12 

0.09 
0.07 
0.07 

No.  1    High 

No.  2    Low                       ' ' 

No.  1    High      " 

No.  2    Low        "             " 

Weight  of  cow  966  lbs. 

DAILY  PHOSPHORUS  METABOLISM   WITH  MILCH  COWS 

EXPERIMENT  III 


Dates 


Total 
P 
fed 


Daily  income  and  outgo 


Fed     Outgo     Feces       Milk     Urine 


Rations 


Total  phosphorus 


Mar.  12-Mar.  18 
Mar.  30-Apr.  5 


26.8 


43.5 
10.9 


14.7 
15.8 


10.17 
0.08 


No.  1    Oat  straw  10  lbs.,  wheat  bran  10 

lbs.,  rice  meal  7  lbs.,  wheat  gluten  Wi 

lbs. 
No.  2    Oat  straw  10  lbs.,  washed  bran 

10  lbs.,  rice  meal  7  lbs.,  wheat  gluten 

2  lbs. 


Nucleo-phosphorus 


Mar.  12-Mar.  18 
Mar.  30-Apr.  5 


77 
16 


23.2 
9.6 


13.4 

9.4 


9.9 
6.1 


3.5 
3.3 


0.00 
0.00 


No.  1 

No.  2 


Soluble  organic  phosphorus 


Mar.  12-Mar.  18 
Mar.  30-Apr.  5 


50.2 
2.6 


1.58 
1.86 


0.53 
0.69 


1.05 
1.17 


0.00 
0.00 


No.  1 

No.  2 


Inorganic  phosphorus 


Mar.  12-Mar.  18 
Mar.  30-Apr.  5 


4.0 
4.2 


53.4 
15.5 


33.1 
4.2 


10.2 
11.2 


10.10 
0.08 


No.  1 
No.  2 


Weight  of  cow  966  lbs. 


310  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Mendel  and  Underhill  (1906) )  conducted  a  series  of  balance  ex- 
periments with  a  female  dog  on  a  diet  of  meat,  cracker  meal  and 
lard.  In  some  periods  sodium  phytate  was  administered  subcutan- 
eously,  and  in  others  the  salt  was  given  by  the  mouth.  The  basal 
ration  was  composed  of  hashed  meat  72  gm.,  cracker  meal  76  gm. 
and  lard  14  gm. ;  the  whole  containing  3.84  gm.  nitrogen,  1.01  gm. 
P205  and  520  calories.      The  numerical  data  are  on  page  311. 

A  part  of  the  authors'  conclusions  are  as  follows : 

"Comparatively  large  doses  of  phospho-organic  acid,  used  as 
the  sodium  salt,  can  be  introduced  into  animals  either  per  os,  subcu- 
taneously,  intraperitoneally,  or  intravenously,  without  unfavorable 
effects.      The  free  acid  is  more  toxic. 

"No  marked  or  immediate  characteristic  effects  of  the  sodium 
salt  upon  general  health  or  nitrogenous  metabolism  have  been  ob- 
served. The  compound  is  readily  absorbed  and  speedily  trans- 
formed within  the  organism.  Its  phosphorus  reappears  in  the  ex- 
creta as  inorganic  phosphates.  No  constant  relation  between  the 
metabolism  of  nitrogen  and  of  phosphorus  was  observed.  In  these 
details  our  experience  with  the  dog  corresponds  with  the  observa- 
tions of  Jordan,  Hart  and  Patten  after  feeding  phytin  to  cattle.  Our 
results  differ  in  showing  that  in  both  the  dog  and  the  rabbit  the  ex- 
cess of  phosphorus  was  almost  entirely  eliminated  through  the  kid- 
neys rather  than  in  the  feces.  This  may  have  an  important  bear- 
ing on  the  possibility  of  producing  laxative  effects  with  phytin. 

"In  our  experimental  animals  purgative  action  could  not  be 
constantly  provoked.  Very  large  doses  were  frequently  effective. 
No  permanent  generalizations  can  be  drawn  from  the  observations 
made  on  this  point." 

The  authors  also  call  attention  to  the  fact  that  the  effects,  with 
their  sodium  salt,  may  differ  from  the  natural  phytin,  because  of  the 
character  of  the  bases  present. 

Horner  (1907)  conducted  balance  experiments  with  a  dog  and 
a  rabbit  which  apparently  showed  that  phytin  may  be  absorbed  and 
retained,  though  the  results  are  not  considered  by  Horner  as  quite 
conclusive. 

McCollum  and  Hart  (1908)  demonstrated  that  calf's  blood  and 
liver  have,  but  muscle  and  kidney  have  not,  the  property  of  cleaving 
phytate  with  the  production  of  inorganic  phosphate.  They  cite 
the  conclusion  of  Scofone  that  the  enzymes  of  the  digestive  tract 
do  not  alter  phytin. 

Hart,  McCollum  and  Humphrey  (1909)  conducted  a  balance  ex- 
periment covering  about  three  and  one-half  months,  with  one  cow, 


PHOSPHORUS  METABOLISM 


311 


<U5  OS  W  O  ^  ^5  ^5  m  Ol  bO  QO  cn  to 

tot-'i-'i-"    •    •    co to to to hrj hr1 
I— 'OocntO'    ■    ;    o~l#>-;-':"J*- 

o'ooooboooboaooo-jJCjncii 
ooooobobobccoaoaoojcjt 


OOtOHCOOCDOJDtDtCQOCO 
SwOO)MM00050tOtO(C 


^ooOTGC--JCo--JGioro--J>-'ao>*>- 


J6OHO00O00MO0 

3  00  CO  -JH  -*J~4 
JOCOCOtO-J~4 


oooooooooopoo 
-coocototoi-T'tototoco^t 


t^h-i-S^^Jcnooco^Ci^-rf^co 


^-4  b>  >&■  oj '*>•  cn  oo  cn  oo  oo  to  <33  to 


|       +1+     +     +++++ 

OOHO^OpOHMNI-H 

io     ;_>bo!-»     ^-a     to^-atococo 


oscocococccoco;s>.co;e.cnccco 
roo^cotocoa:cocococoo^co 


tOtOI-'tOl-'IOJ-'COJ-'COCOJ-'h-' 

eoeociton-ccoih-cDcocncoco 


OiOCJicocotocoococopo-JOj 


c  n  o 

O'er  3 

o  o  c->- 

S  C  3 

p  i°  rr 

n>  <t>  2 

C  C  o 
oi  oi  p. 

S3.' 

re'o" 

0  o 

<->■  J+ 

o'S' 
3  B 

aw 

p  p 

01  ►?) 

£° 

OO 

COCO 
COCO 

BB 


coco 
o*B  a-3 
O  c+  O  rf 
C  1  B  i 

re  2  re 
o  2.  o 
3  o  B  w 
oi  3.01  g^ 


3  o  (B  (S  (1  O 
o*3  p.p.p.3 

H  >d   01  01   01  W 

Sjppfiit 

o  S.SS5H, 
C  o -•-•-•  5 

QOOOO 
05OT 
Oq  «q  3 

bbp 


E5  s  « 

S  p  p 

W  oi  oi 

O  EL  EL 


B     3     B 


OO; 

1 5 

3.8 

S  p 


3S1 

oo 


hj     *t    $        P* 
O     O     O 


W      I  W 


w 


> 
W 

o 
f 


» 

i— i 

3 

H 
H 

02 


a 


o 

f 

Q 
» 
3 


312 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


in  a  study  of  phytin  metabolism.  The  phytin  was  as  present  in 
wheat  bran,  and  as  prepared  from  the  same  as  a  crude  potassium 
salt.  Whole  bran  was  compared  with  washed  bran,  and  with  washed 
bran  plus  potassium  phytate.  The  washing  process  removes,  of 
course,  a  great  variety  of  compounds  other  than  the  one  of  especial 
interest. 

These  authors  reached  the  conclusion  that  the  phytin  of  wheat 
bran  does  not  have  a  specific  effect  on  the  production  of  milk,  or  on 
the  production  of  fat  in  the  milk,  and  that  previous  results,  appar- 
ently showing  such  effects,  have  been  due  to  the  individuality  of  the 
subjects.  Phytin  was  found  to  be  diuretic  because  of  its  potassium 
content,  and  also  laxative  because  of  the  elimination  of  phosphorus 
and  accompanying  bases  by  way  of  the  feces. 

Cook  (1909)  conducted  a  metabolism  and  tissue  analysis  ex- 
periment with  six  rabbits,  in  the  comparison  of  phytin  with  sodium 
phosphates,  when  taken  in  excess  of  the  maintenance  requirement. 
Two  rabbits  were  fed  on  corn,  oats,  and  vegetables  as  controls ;  the 
remaining  four  received  a  ration  of  carrots,  wheat  gluten,  starch, 
sugar,  olive  oil  and  salt  mixture,  two  receiving  phytin  in  addition, 
and  the  other  two  inorganic  phosphorus  in  the  shape  of  sodium 
dihydrogen  phosphate  and  disodium  hydrogen  phosphate.  The 
phytin  was  prepared  from  wheat  bran.  The  condensed  nitrogen 
and  phosphorus  balance  data  are  as  follows : 

AVERAGE   DAILY   NITROGEN   AND   PHOSPHORUS    (P206)    BALANCES 

WITH  RABBITS  COMPARING  PHYTIN  AND  SODIUM 

PHOSPHATES— Grams 


Dates 

Rabbit 

No. 

Weight, 
averag-e 

Nitrogen 

Phosphorus    P20s^ 

Intake 

Balance 

Intake 

Balance 

Form  of 
phosphorus 

Nov 

17-Feb.  15.... 

1 

2 
3 
4 

1618 
1645 
1585 
1981 

1.154 

1.353 

"  1.520 

1.396 

+0.23 
+0.32 
+0.38 
+0.33 

0.381 
0.415 
0.501 
0.459 

+0.14 
+0.14 
+0.18 
+0.18 

Phytin 
Phytin 
Phosphate 
Phosphate 

Feb. 

17-Mar.  15.... 

1 
3 

4 

1550 
1796 
1994 

1.479 
1.747 
1.485 

+0.26 
+0.40 
+0.18 

0.456 
0.541 
0.498 

+0.166 
+0.166 
+0.156 

Phytin 

Phosphate 

Phosphate 

These  data  do  not  show  unmistakable  differences  in  the  value 
of  these  compounds  under  the  conditions  of  these  experiments.  It 
is  worthy  of  note  that  the  calcium  and  magnesium  balances  were 
positive.  A  part  of  the  inconclusive  character  of  the  results  must 
be  due  to  the  fact  that  there  was  loss  of  live  weight  in  5  out  of  the 
7  periods. 

The  tissue  analyses  revealed  more  decided  differences,  though 
their  significance  is  as  yet  unexplained.      The  autopsy  revealed  ab- 


PHOSPHORUS  METABOLISM 


313 


normal  conditions  in  the  livers,  kidneys  and  lungs  of  the  rabbits 
which  received  the  phosphorus  compounds,  the  livers  all  being  pale 
in  color,  and  enlarged.  The  livers  of  the  rabbits  which  had 
received  phytin  showed  marked  fatty  degeneration,  as  also 
did  one  of  those  receiving  inorganic  phosphates,  though  in  a 
less  pronounced  way.  The  kidneys  of  the  inorganic  phosphate  rab- 
bits showed  parenchymatous  degeneration.  Below  are  analytical 
data  from  several  tissues  of  these  rabbits. 

CHEMICAL  ANALYSES  OF  BODIES  OF  RABBITS  AS  AFFECTED  BY 
PHYTIN  AND  INORGANIC  PHOSPHATE— Dry  Basis— Percent 


Nitro- 

Ash 

Cal- 

Magne- 

Ether 

Phosphoric 

acid 

Ether- 

Ether -alco- 

gen 

cium 

sium 

extract 

Total 

alcohol- 
soluble 

hol-soluble 
in  terms 
of  total 

Bones 

4.53 

55.56 

8.86 

0.22 

11.33 

23.96 

0.055 

0.23 

Livers.  . . 
Blood 

7.64 
14.72 

3.78 
5.64 

0.00 
Trace 

0.00 
0.00 

44.95 
Trace 

1.98 
0.75 

0.680 
0.008 

34.34 
1.07 

Mean  of  rabbits  1  and 
2  fed  organic  phos- 
phorus. 

Brains. . . 

6.12 

7.00 

0.31 

0.09 

43.23 

3.96 

2.350 

59.34 

Nerves. . . 

5.52 

6.54 

0.21 

0.11 

46.96 

3.72 

2.390 

64.26 

Teeth.... 

74.65 

24.73 

1.35 

35.31 

Bones 

4.68 

55.92 

7.74 

0.15 

9.39 

26.33 

0.061 

0.23 

Livers.  . . 
Blood.... 

9.41 
14.42 

4.73 
4.72 

0.00 
Trace 

0.00 
0.00 

34.48 
Trace 

2.56 
0.69 

0.854 
0.008 

33.36 
1.16 

Mean  of  rabbits  3  and 
4  fed  inorganic  phos- 
phorus. 

Brains. . . 

6.52 

7.17 

0.27 

0.06 

43.89 

4.07 

1.160 

28.51 

Nerves. . . 

3.72 

5.83 

0.28 

0.06 

47.34 

4.28 

1.470 

34.35 

Teeth.... 

76.10 

24.65 

1.22 

34.70 

Bones 

4.32 

57.47 

10.17 

0.23 

5.12 

25.93 

0.069 

0.27 

Livers. . . 

11.90 

5.67 

0.00 

0.00 

14.47 

2.85 

1.090 

38.24 

Blood.... 

14.42 

4.55 

0.44 

0.19 

Trace 

0.58 

0.037 

6.33 

Mean  of  rabbits  5  and 

Brains. . . 

6.85 

7.82 

0.51 

0.08 

38.50 

4.16 

1.750 

42.07 

6  normally  fed. 

Nerves. . . 

4.39 

5.97 

0.44 

0.12 

44.34 

3.90 

2.310 

59.23 

Teeth.. . . 

75.17 

28.35 

1.15 

35.63 

Among  the  more  prominent  results  here  set  forth  are:  (1)  the 
low  calcium  content  of  the  tissues,  (2)  the  increase  above  normal 
of  the  alcohol-ether  soluble  phosphorus  of  the  brain  of  the  rabbits 
which  received  phytin,  and  (3)  the  decrease  below  normal  of  the  al- 
cohol-ether soluble  phosphorus  of  the  brain  and  nerves  of  the  rab- 
bits which  received  inorganic  phosphates. 

Tyshnjenko  (1909)  found,  in  balance  experiments  with  5  labor- 
atory assistants  on  a  diet  of  bread,  meat,  milk  and  butter,  that  the 
addition  of  sodium  glycerophosphate  caused  a  loss  of  phosphorus 
from  the  body,  while  phytin  led  to  retention.  We  have  seen  only 
Maly's  abstract  of  this  article  of  117  pages. 

Starkenstein  (1910)  states  that  the  organs  of  infants  contain 
larger  amounts  of  inosite  than  those  of  adults,  and  that  it  originates 
in  the  inosite-phosphoric  acid  of  the  food,  which  adults  are  able,  in 
larger  part  than  infants,  to  decompose,  the  remainder  passing  off 


314  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

unchanged  in  the  urine.  Starkenstein,  therefore,  regards  inosite 
as  of  no  physiological  importance  in  the  animal  body,  but  as  a  waste 
product  of  phosphoric  acid  metabolism;  and  he  considers  inosite- 
phosphoric  acid  as  of  importance  only  as  a  source  of  phosphorus, 
and  also  of  value  in  certain  pathological  conditions  of  the  osseous 
system.  We  shall  stand  in  a  fair  way  to  clear  up  these  problems 
when  we  have  discovered  a  satisfactory  method  for  the  estimation 
of  inosite-phosphoric  acid. 

Rogozinski  (1910)  studied  phosphorus  metabolism  in  an  adult 
dog  in  7  continuous  5-day  experiments,  without  intervals  between 
collection  periods.  Phytin,  lecithin  and  sodium  phosphate  were  ad- 
ded, in  different  periods,  to  a  diet  of  meat,  rice  and  lard.  Rogozinski 
found  no  pronounced  influence  of  these  compounds  on  the  nitrogen 
excretion  in  the  urine,  when  they  were  added  to  a  ration  sufficient  to 
maintain  the  animal. 

The  phosphorus  of  the  sodium  phosphate  appeared  quantita- 
tively in  the  urine.  The  total  lecithin  fed  to  the  dog  was  split,  and 
the  phosphorus  excreted  in  the  urine  as  inorganic  phosphate;  the 
phosphorus  of  phytin  passed  into  the  urine  to  the  extent  of  about 
30  percent  of  the  total,  the  remainder  being  excreted  in  the  feces, 
where  it  could  be  determined  as  such ;  the  nitrogen  and  phosphorus 
balances  were  found  to  a  high  degree  independent  of  each  other. 

In  a  15-day  experiment  on  himself  Rogozinski  added  2  gm.  of 
phytin  to  a  mixed  diet  (which  was  not  analyzed)  on  the  6th  to  10th 
days.  The  phytin  was  completely  split  in  the  digestive  tract;  a 
small  part  of  the  phytin  phosphorus  was  retained,  and  the  rest  was 
excreted  as  inorganic  phosphorus  in  the  feces ;  the  feces  contained 
an  abundance  of  lecithin-like  compounds;  no  inosite  was  found  in 
the  urine  after  feeding  phytin;  the  bacteria  of  human  feces  can 
split  off  inorganic  phosphoric  acid  from  phytin. 

Donath  (1911)  reports  phytin  as  a  powerful  stimulant  to  the 
appetite,  and  states  that,  in  dogs  having  Pawlow  fistulae,  it  increas- 
es the  flow  of  gastric  juice. 

Sodium  phytate  in  large  quantities  was  found  toxic  to  rabbits 
by  A.  R.  Rose  (1911).  It  appears  that  1.7  gm.  per  kilogram  of  body 
weight  is  fatal.  Rose  (1912a)  has  conducted  an  extensive  study 
on  the  metabolism  of  phytin  with  a  milch  cow.  Organic  as  well  as  in- 
organic phosphorus  of  the  food  was  eliminated  very  largely  in  the 
form  of  inorganic  phosphorus  in  the  feces,  the  amount  of  phospho- 
rus in  the  urine  being  very  small.  The  addition  of  calcium  phytate 
increased  the  potassium,  both  in  the  urine  and  feces,  and  changed 
the  path  of  elimination  of  part  of  the  magnesium  from  the  kidney  to 
the  intestine.      The  calcium  added  as  calcium  phytate  was  almost 


PHOSPHORUS  METABOLISM  315 

entirely  eliminated  by  the  intestine  immediately  after  administra- 
tion. The  calcium  of  the  urine  increased  with  decreasing  phos- 
phorus in  the  rations,  and  decreased  when  calcium  phytate  was 
added.  The  volume  of  the  milk  fluctuated  inversely,  and  the  total 
amount  of  fat  in  the  milk  directly,  as  the  amount  of  phytin  phospho- 
rus in  the  rations.  The  increase  of  milk  flow  on  removal  of  the 
phytin  was  not  a  mere  dilution.  Except  for  the  change  (increase) 
in  the  amount  of  fat,  the  composition  of  the  milk  was  not  material- 
ly altered. 

These  results  sustain  the  earlier  work  of  Jordan,  Hart  and  Pat- 
ten, but  since  the  cow  used  by  Rose  was  one  of  the  same  that 
were  used  in  the  work  of  Jordan,  Hart  and  Patten,  the  factor 
of  individuality  was  not  excluded. 

G.  di  Gregorio  (1912)  concluded  that  the  administration  of  1-2 
gm.  daily  of  phytin  can  check  a  pathological  phosphaturia,  and  un- 
der normal  conditions  can  diminish  loss  of  phosphorus. 

Santonocetto  (1912)  studied  nitrogen  metabolism  as  influenced 
by  the  administration  of  phytin.  He  concluded  that  under  the  in- 
fluence of  phytin  the  breaking-down  of  the  cells  is  retarded,  and  the 
absorption  of  food  nitrogen  is  promoted,  the  fecal  nitrogen  being 
reduced  30-75  percent,  and  the  urine  nitrogen  7.12  percent. 

Venturi  and  Masella  (1913)  studied  the  influence  of  phytin  in- 
gestion on  the  balance  of  nitrogen,  and  on  the  qualitative  distribu- 
tion of  the  nitrogenous  metabolites.  The  experimental  subject  was 
one  of  the  authors.  After  a  preliminary  period  of  4  days  phytin 
was  administered  in  the  amount  of  1.5  gm.  daily  for  5  days;  then 
followed  an  after-period  of  2  days,  and  a  second  phytin  period  of  5 
days,  during  which  2.5  gm.  of  the  compound  was  fed  daily.  The 
daily  data  show,  in  the  phytin  periods  as  compared  with  the  fore- 
and  after-periods,  a  marked  decrease  of  urinary  nitrogen,  especially 
in  the  form  of  urea,  creatinin,  hippuric  acid  and  xanthin  bases,  the 
uric  acid  and  ammonia  remaining  without  great  change.  The  feces 
nitrogen  was  decreased,  the  nitrogen  retention  being  prominently 
increased.  The  authors  note  the  agreement  of  their  results  with 
those  of  Santonocetto.  See  also  Forbes  and  associates  (1914),  Nutr. 
Val.  Org.  and  Inorg.  P. 

CLINICAL  EXPERIMENTS  WITH  PHYTIN 

Secheret  (1904)  made  a  historical  and  critical  study  of  phytin 
therapy,  with  some  observations  on  animals  after  the  introduction 
of  phytin,  and  also  of  61  human  patients  under  treatment  with  phy- 
tin. Most  of  the  animals  receiving  phytin,  otherwise  than  by  the 
mouth,  died  soon.      Hypodermic  injections    were,    therefore,    not 


316  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

recommended.  Dosage  was  discussed,  and  phytin  recommended  in 
a  great  number  of  pathological  states,  especially  as  a  general  tonic, 
and  to  increase  the  blood  corpuscles,  in  constitutional  debility,  from 
whatever  cause.  Phytin  is  said  to  possess  a  very  active  pharmaco- 
dynamic action,  exciting  the  internal  nutrition  Of  the  tissues  and 
cells,  and  increasing  the  nitrogen  elimination. 

Bardet  (1905)  has  given  study  to  the  condition  of  general 
"demineralization,"  which  may  be  due  to  any  such  illness  of 
long  standing  as  results  in  a  disturbance  of  the  normal  balance  be- 
tween assimilation  and  losses.  Of  17  cases  examined  4  are  re- 
ported ;  one  a  woman  23  years  old,  in  a  low  state  because  of  a  long 
siege  of  typhoid  fever;  another  was  a  wet-nurse,  aged  28,  who  was 
in  a  debilitated  state ;  a  third  was  a  boy  16  years  old  who  had  be- 
come highly  neurasthenic  through  overstudy;  and  a  fourth  was  a 
rapidly  growing  boy  of  14  years,  who  was  in  a  much  disturbed  state. 
Adrian's  extract  of  cereals  (containing  4  percent  P205,  and  consid- 
erable potassium,  magnesium,  and  some  manganese,  all  in  organic 
combination,  largely  as  a  phytin)  was  administered  with  sodium 
methyl  arsenate.  Fifteen  to  twenty-five  grams  of  the  extract  were 
given  in  solution  each  day  for  30  or  40  days,  which  treatment 
brought  about  almost  complete  restoration  to  normal  condition. 
Since  this  cereal  extract  contained  in  assimilable  form  nutrients  of 
which  the  body  stood  in  need,  it  was  probably  of  some  value,  though 
the  use  of  arsenic  adds  a  factor  to  the  problem  which  makes  impos- 
sible a  positive  interpretation  of  the  results. 

Novi  (1908)  submitted  data  apparently  showing  that  the  ad- 
ministration of  phytin,  1-1.5  gm.  per  day,  decreased  the  urinary 
phosphorus  in  antirabes  treatment  from  69.11  to  46.07  percent  of 
the  intake.      Feces  figures  were  not  submitted. 

Novi  (1909)  reported  that  in  consequence  of  the  injection  of  1- 
percent  water  solution  of  phytin  into  the  dorsal  lymph  sack  of  the 
frog  the  muscle  power  was  2-4  times  higher  than  normal.  Calcium 
chloride  produced  no  such  effect,  but  a  mixture  of  sodium,  mag- 
nesium and  calcium  chlorides  produced  the  same  effect  as  the  phy- 
tin, as  also  did  magnesium  chloride  alone.  Sodium  glycerophos- 
phate produced  no  effect,  but  calcium  glycerophosphate  doubled  the 
muscular  power. 

Favorable  results  from  the  use  of  phytin,  by  ingestion,  in  cases, 
of  gastric  ulcer  are  reported  by  Wolpe  (1911). 

Other  clinical  reports  from  the  use  of  phytin  in  human  medical 
practice  are  those  of  Loewenheim  (1904),  Gilbert  and  Lippmann 


PHOSPHORUS  METABOLISM  317 

(1904),  Fiirst  (1904),  Gianasso  and  Ovazza  (1905),  Winterberg 
(1905),  Dambre  (1905),  Wechsler  (1905),  Maestro  (1905a),  and 
Weissmann  (1908).  The  results  of  these  studies  have  been  almost 
uniformly  favorable,  as  shown  by  improved  state  of  nutrition  gen- 
erally, increased  appetite,  improved  blood  conditions,  etc.  Emphasis 
is  placed  on  the  fact  of  the  very  high  phosphorus  content  of  this 
compound,  making'  it  a  superior  vehicle  for  the  administration 
of  phosphorus  in  an  organic  form. 

Summary.  Conservatism  in  the  acceptance  of  results  in  the 
clinical  study  of  phytin  is  especially  to  be  recommended  because  of 
the  abundance  of  phytin  and  related  compounds  in  some  of  our  com- 
mon foods,  which  produce  no  marked  specific  symptoms.  On  the 
other  hand,  we  are  obliged  also  to  admit  the  possibility  that  this 
compound  as  administered  in  an  uncombined  state  may  have  a  dif- 
ferent effect  and  method  of  usefulness  from  the  same  as  normally 
combined  in  foodstuffs.  There  is  not  the  same  ground  for  this 
latter  hypothesis,  however,  that  there  is  in  the  case  of  lecithin,  since 
phytin  even  though  absorbed  as  such,  unsplit,  must  be  broken  up  be- 
fore it  can  be  of  use  for  constructive  purposes,  phytin  not  being  a 
constituent  of  the  animal  body. 

A  measure  of  uncertainty  must  exist  in  regard  to  results  in  this 
field  until  an  understanding  of  the  chemistry  of  phytin  and  related 
compounds  has  been  attained.  It  seems  quite  probable  that  the 
name  phytin  has  been  associated  with  a  variety  of  compounds. 
There  is  no  question,  however,  as  to  the  absorbability  of  phytin,  nor 
of  its  usefulness  in  animal  metabolism. 

Phytin  is  readily  soluble  in  the  hydrochloric  acid  of  the  gastric 
juice  and  is  mostly  absorbed  without  decomposition  by  the  digestive 
enzymes.  It  is  decomposed,  however,  before  utilizaton  by  the  tis- 
sues, and  to  such  extent  as  it  is  not  retained,  its  phosphorus  is  elim- 
inated in  the  urine  and  feces.  In  cattle  and  in  human  beings  phytin 
phosphorus  is  eliminated  as  inorganic  phosphates,  following  the 
usual  course  as  determined  by  the  nature  of  the  diet  and  by  the  in- 
organic bases  present.  In  dogs,  however,  a  considerable  part  of 
the  phytin  phosphorus  seems  to  be  eliminated  in  the  feces,  where  it 
is  found  present  as  phytin. 

The  power  to  split  phytin  has  been  found  to  reside  in  the 
blood  and  liver,  but  not  in  the  muscle  and  kidney  of  the  calf.  It 
may  also  be  decomposed  by  intestinal  bacteria. 

The  contribution  of  phytin  to  the  laxative  and  diuretic  effects 
of  rations  is  through  the  inorganic  bases  contained.  There  has 
not  yet  been  established  a  definite  relation  between  phytin  ingestion 
and  nitrogen  metabolism. 


318  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

We  are  not  yet  able  to  say  whether  or  not  phytin  has  specific 
effects  in  animal  nutrition,  but  there  is  certainly  nothing  strikingly 
characteristic  about  the  method  of  its  use.  Apparently,  at  least 
its  chief  superiorities  as  a  phosphorus-carrying  nutrient  lie  in  its 
ready  solubility  and  in  its  high  phosphorus  content. 

THE  NUTRITIVE  VALUES  OF  ORGANIC  AND  INORGANIC 

PHOSPHORUS 

METABOLISM  EXPERIMENTS  WITH  MEN  AND  ANIMALS 

Considering  the  phosphorus  of  the  animal  body  as  a  whole  the 
most  obvious  distinction  among  the  various  groups  of  its  compounds 
is  that  certain  of  these  are  organically  combined,  as  a  part  of  the  liv- 
ing tissue,  being  fundamentally  involved  in  all  vital  activities,  while 
others  are  simple  salts  of  the  mineral  bases  either  deposited  in  sup- 
porting structures  or  dissolved  in  the  body  fluids. 

In  both  cases  the  phosphorus  itself,  so  far  as  known,  is  present 
in  the  same  completely  oxidized  form  as  phosphoric  acid,  differing 
only  in  its  chemical  relationships.  This  discussion  is  a  considera- 
tion of  the  evidence  as  to  the  nutritive  limitations  imposed  by  these 
differences  in  relationship  of  phosphoric  acid.  We  wish  to  know 
whether  organic  and  inorganic  phosphorus  compounds  can  serve, 
equally  well,  all  the  requirements  of  the  body  for  phosphorus  under 
all  conditions  of  life. 

Let  us  consider  first  the  eggs  of  birds,  for  the  egg  assuredly 
contains  all  of  the  nutrients  required  by  the  fully  formed  animal. 
Examining  the  phosphorus  of  eggs  we  find  a  wealth  of  organic  com- 
pounds but,  at  the  most,  mere  traces  of  inorganic  phosphates.  It 
is  thus  obvious  that  organic  phosphorus  compounds  can  serve  all  of 
the  needs  of  the  body  for  this  element.  But  our  interest  is  in  food- 
stuffs ;  let  us  consider  the  first  natural  food  of  young  mammals,  the 
mother's  milk.  Are  the  comparative  amounts  of  organic  and  inor- 
ganic phosphorus  related  to  the  food  requirements  of  the  young  ani- 
mal, or  do  they  simply  represent  their  relative  availability  for  milk 
production  in  the  maternal  organism  ?  However  this  may  be,  if  there 
is  not  an  adaptation  of  the  character  of  the  food  to  the  requirements 
of  the  young,  then  there  must  be  an  adaptation  of  the  method  of  de- 
velopment of  the  young  to  the  possibilities  of  the  food,  in  either  case 
a  harmony  of  objects  to  be  attained  and  means  for  their  attainment. 

In  this  light  we  would  naturally  assume  that  both  organic  and 
inorganic  phosphorus  compounds  are  of  benefit  to  the  animal;  for 


PHOSPHOEUS  METABOLISM  319 

both  classes  are  represented  in  milk  by  several  individuals  each,  and 
in  the  whole  literature  of  the  subject  there  is  scarcely  a  dissenting 
voice  raised  against  this  idea.  Both  organic  and  inorganic  phos- 
phorus are  absorbed  and  retained. 

But  now  we  come  to  the  parting  of  the  ways.  If  organic  phos- 
phorus can  serve  all  of  the  requirements  of  the  body  for  phosphorus, 
and  if  inorganic  phosphorus  can  be  absorbed  and  retained,  are  or- 
ganic and  inorganic  phosphorus  equally  useful  for  all  of  the  purpos- 
es for  which  the  body  needs  phosphorus  ?  Our  economic  reason  for 
desiring  an  answer  to  this  question  lies  in  the  relative  accessibility 
of  organic  and  inorganic  phosphorus  for  use  as  food.  If  rock  phos- 
phate and  old  bones  can  furnish  us  phosphorus  in  the  forms  most 
advantageous  for  the  growth  of  animals,  we  are  wasting  much 
money  on  milk,  eggs  and  beef;  for  there  are  much  cheaper  sources 
than  these,  of  protein,  fat  and  carbohydrates. 

In  this  consideration  let  us  bear  in  mind  that  our  interest  as  ag- 
ricultural scientists  is  not  so  much  in  bare  physiological  minima  as 
in  maximum  practical  optima,  for  the  whole  range  of  success  and 
profit  in  animal  production  lies  close  to  the  latter. 

It  would  seem  that  so  simple  a  problem  ought  readily  to  be 
solved,  but  when  we  approach  the  subject  by  direct  experiment  we 
find  the  course  beset  with  hazards,  and  there  never  has  been 
unanimity  of  opinion  as  to  the  facts.  Among  those  circumstances 
which  have  contributed  to  this  difference  in  opinion  are: 

(1)  An  inclination  to  ascribe  to  all  animals  under  all  condi- 
tions of  life  all  capacities  of  synthesis  possessed  by  any  animal  un- 
der any  condition,  natural  or  experimental,  no  matter  how  great 
the  stress  of  attending  circumstance. 

(2)  Drawing  conclusions  from  very  short  balance  experi- 
ments, without  due  regard  to  states  and  habits  of  nutrition  as  de- 
termined by  previous  feeding. 

(3)  A  failure  to  distinguish  between  physiological  minima 
and  practical  optima. 

(4)  Drawing  conclusions  from  mere  gain  in  weight,  without 
actual  estimation  of  the  compounds  of  interest  or  measurements  of 
functional  efficiency  in  the  experimental  animals  and  in  carefully 
selected  controls,  the  error  in  so  doing  being  that  it  implies  the 
maintenance  of  constancy  of  composition  and  function,  the  variabil- 
ity of  which,  as  affected  by  feeding,  is  usually  underestimated. 

(5)  Drawing  conclusions  from  analyses  of  parts  of  animals, 
a  complete  chemical  accounting  being  desirable. 


320  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

(6)  Failure  fairly  to  meet  the  great  difficulties  of  compound- 
ing rations  which  do  not  differ  in  essential  ways  other  than  in  the 
point  of  interest,  that  is,  in  the  condition  in  which  the  phosphorus  is 
present. 

(7)  It  is  quite  possible  that  useful  enzymes  associated  with 
the  organic  phosphorus  compounds  have  exerted  a  determining  in- 
fluence on  the  results. 

(8)  Unsatisfactory  mineral  salt  accompaniments  may  have 
affected  the  usefulness  of  the  phosphorus  compounds  in  such  ways 
as  to  result  in  unfair  comparisons. 

(9)  The  specific  physiological  action  of  isolated  compounds 
of  phosphorus,  as  used  in  nutrition  investigations,  undoubtedly  dif- 
fers from  the  effects  of  the  same  compounds  in  their  natural  rela- 
tionships in  foodstuffs. 

We  shall  review,  in  brief,  the  evidence  on  the  subject. 

W.  S.  Hall  (1896),  from  feeding  experiments  with  mice,  came  to 
the  conclusion  that  the  salts  organically  combined  with  casein  have 
a  value  greater  than  salts  not  so  combined. 

Steinitz  (1898)  conducted  balance  experiments  on  dogs  to  com- 
pare organic  and  inorganic  phosphorus  compounds  as  nutrients. 
Nutrose,  a  sodium-calcium-casein  compound,  vitellin,  and  myosin 
prepared  from  horse  flesh  were  each  added,  in  different  periods,  to  a 
basal  ration  of  bacon,  rice  starch  and  mineral  salts ;  the  casein  prep- 
aration and  the  vitellin  both  containing  organic  phosphorus,  and  the 
myosin  being  at  least  practically  free  from  organic  phosphorus. 
Phosphorus  was  stored  abundantly  on  the  organic  phosphorus  ra- 
tions, but  the  retention  was  almost  nothing  on  the  myosin  ration, 
though  in  all  cases,  including  the  myosin  ration,  nitrogen  was  stored 
in  considerable  quantity.  The  salt  mixture  used  with  the  organic 
phosphorus  compounds  contained  chlorides  and  citrates  only,  while 
the  salts  used  with  the  myosin  were  calcium,  magnesium  and  potas- 
sium phosphates,  sodium  chloride  and  ferric  citrate.  The  experi- 
mental periods  were  5-9  days  each,  following  8-day  fasting  periods. 

Rohmann  (1898)  found,  in  balance  experiments  on  a  dog,  with 
a  diet  of  lard,  rice  starch,  salt  and  either  a  phosphorus-containing 
protein,  or  a  phosphorus-free  protein  and  a  phosphate,  that  both 
nitrogen  and  phosphorus  retention  were  favored  by  the  phosphorus- 
containing  protein  much  more  than  by  the  phosphorus-free  protein 
and  phosphate.  With  nutrose  and  vitellin  the  nitrogen  retention 
was  31.0  and  42.2  percent,  respectively,  of  the  intake;  while  with 
myosin  and  edestin  the  retention  was  but  0.10  and  0.11  percent  of 
the  intake.  Likewise  with  the  nutrose  and  vitellin  the  phosphorus 
retention  was  8.8  and  20.9  mg.  per  day,  while  with  myosin  and  edes- 
tin the  retention  was  0.1  mg.  per  day  in  both  cases. 


PHOSPHORUS  METABOLISM  321 

Leipziger  (1899)  studied  metabolism  in  a  dog  on  a  ration  which 
was  very  low  in. organic  phosphorus.  The  ration  was  composed  of 
edestin,  fat,  starch,  salt,  beef  extract  and  water.  The  dog  was 
fasted  for  6  and  10  days,  respectively,  in  the  fore-periods  of  two 
tests,  and  then  fed  for  6  days  in  each  of  the  two  main  periods. 

The  phosphorus  intake  in  the  two  experiments  was  1.874  and 
2.022  gm.  per  day,  of  which  0.016  and  0.022  gm.  was  organic.  The 
dog  retained  0.0078  and  0.095  gm.  of  phosphorus  per  day  in  these 
tests,  the  nitrogen  and  calcium  balances  also  being  positive.  The 
subject  of  these  experiments  was  the  dog  referred  to  as  "Dog  II"  in 
Steinitz's  experiments. 

Leipziger  considered  that  his  data  did  not  afford  evidence  as  to 
the  condition  in  which  the  phosphorus  was  retained,  but  thought 
that  synthesis  of  phosphorus-containing  protein  from  phosphorus,- 
f  ree  protein  and  phosphates  was  improbable ;  also  that  the  phospho- 
rus retention  was  less  than  it  would  have  been  on  a  ration  contain- 
ing more  phosphorized  protein.  The  organic  phosphorus  content 
of  this  ration  was  sufficient  to  render  impossible  the  solution  of  the 
problem  of  phosphorized  protein  synthesis  except  by  negative  re- 
sults— which  were  not  obtained. 

Zadik  (1899)  conducted  nitrogen  and  phosphorus  balance  ex- 
periments with  dogs  in  the  comparison  of  phosphorized  proteins 
with  phosphorus-free  proteins  and  inorganic  phosphates.  The 
compounds  of  interest  were  casein,  vitellin  and  edestin.  The  basal 
ration  was  composed  of  starch,  bacon,  sodium  citrate  or  carbonate, 
and  a  salt  mixture  of  phosphates,  chlorides,  magnesium  citrate  and 
sugar.  The  numerical  data  contain  numerous  inconsistencies,  on 
account  of  which  we  do  not  transcribe  them,  but  the  errors  seem 
not  to  be  of  a  degree  to  modify  at  all  the  significance  of  the  results. 
With  casein  or  vitellin  there  was  marked  retention  of  phosphorus; 
with  edestin  and  disodium  phosphate  there  was  always  loss. 

Zadik  concluded  that  the  animal  organism  does  not  have  the 
power  to  build  from  phosphorus-free  proteins  and  phosphates  the 
organic  phosphorus  compounds  necessary  for  the  life  of  the  cells. 
The  organic  phosphorus  of  casein  and  vitellin  was,  under  his  experi- 
mental conditions,  at  least,  more  useful  than  the  inorganic  phos- 
phates ;  also,  the  phosphorus  of  vitellin  was  stored  in  larger  propor- 
tion to  the  intake  than  was  the  phosphorus  of  casein. 

Ehrlich  (1900)  conducted  five  balance  experiments  with  dogs  in 
a  comparison  of  phosphorized  proteins  with  phosphorus-free  pro- 
tein plus  inorganic  phosphates.  The  results  tend  to  show  that 
the  phosphorized  proteins,  casein  and  vitellin,  have  a  greater  useful- 


322 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


ness  to  the  animal,  in  the  sense  of  favoring-  phosphorus  retention, 
than  does  the  phosphorus-free  edestin  plus  inorganic  phosphates. 
The  periods  were  3-7  days  in  length.  Certain  unfavorable  condi- 
tions render  the  results  of  doubtful  value. 

Kornauth  (1900)  compared  synthetic  "nucleins,"  prepared 
after  the  method  of  Liebermann,  by  the  precipitation  of  egg  albu- 
min and  blood  serum  albumin  with  metaphosphoric  acid,  with  natur- 
al proteins  in  the  form  of  meat,  skin,  aleuronate  (a  vegetable  casein) , 
casein  and  conglutin  (from  lupines) .  The  results  on  maintenance 
of  nitrogen  equilibrium  hardly  warrant  conclusions,  but  the  evidence 
is  quite  satisfactory  in  showing  that,  for  maintenance  of  phosphorus 
equilibrium,  very  much  more  phosphorus  in  the  so-called  synthetic 
nucleins  is  necessary  than  of  phosphorus  in  the  natural  proteins. 

Steinitz  (1900)  reports  several  attempts  to  rear  young  dogs 
on  artificial  food  mixtures  containing  their  protein  as  casein, 
edestin,  milk,  nucleoprotein  from  calf  liver,  and  vitellin.  Other 
foods  used  were  rice  starch,  lard,  bacon  and  inorganic  salts.  The 
experiments  were  mostly  of  short  duration,  on  account  of  unfavor- 
able termination.  None  were  carried  through  to  a  satisfactory 
demonstration  of  the  sufficiency  of  the  diet. 

Gottstein  (1901)  conducted  a  metabolism  experiment  with  a 
dog  in  two  five-day  balance  periods  in  which  casein  was  opposed  to 
edestin.      The  results  were  inconclusive. 

Ehrstrom  (1903a)  conducted  an  experiment  which  bears  on  the 
relative  value  of  organic  and  inorganic  phosphorus.  The  condensed 
data  are  as  below. 


DAILY  NITROGEN  AND  PHOSPHORUS  BALANCES   WITH  A   GROWN 

MAN  RECEIVING  ORGANIC  AND  INORGANIC  PHOSPHORUS 

COMPOUNDS— Grams 


Length  of  balance 
period 

Diet 

N 
Intake 

N 
Balance 

P 

Intake 

P 
Balance 

Freely  chosen  diet  of  ordinary  foods 

Proton-bread  and  milk 

Ordinary  white  bread,  milk  and  CaHP04 

+17.37 
+17.86 
+12.55 

-1.33 

+0.58 
-1.81 

+2.476 
+2.090 
+2.271 

+0.610 

+0.642 

+0.227 

Less  phosphorus  was  stored  when  a  certain  portion  of  the  total 
was  administered  as  dibasic  calcium  phosphate  than  when  the  same 
amount  was  taken  as  proton — a  casein  preparation ;  in  these  last  two 
periods,  then,  we  have  opposed  to  each  other  a  typical  phosphopro- 
tein  and  an  inorganic  phosphate,  added  in  each  case  to  a  bread  and 
milk  ration.  The  author  does  not  submit  data  showing  that  the  phos- 
phate ration  contained    bases,    especially    calcium,    sufficient    in 


PHOSPHORUS  METABOLISM 


323 


amount,  relative  to  acid  elements,  to  render  conditions  as  favorable 
for  phosphorus  storage  as  in  the  proton  ration.  The  periods  also 
are  very  short  for  mineral  metabolism  work.  The  data  seem  to 
show  the  organic  phosphorus  the  more  useful. 

Hirschler  and  Terray  (1902,  1905)  compared  bone  dust  with 
eggs  as  sources  of  phosphorus  for  growing  dogs.  A  portion  of  the 
data  are  below. 


METABOLISM  EXPERIMENTS  ON  DOGS  WITH  ORGANIC  AND 

INORGANIC  PHOSPHORUS  COMPOUNDS 

Daily  Amounts — Grams 


Gain  or 

Intake 

Phosphorus 

Daily  balances 

Period 

loss  in 

and 

weight  of 

Diet 

days 

dog- 
Kg. 

N 

P2O5 

CaO 

Urine 

Feces 

N 

P2O5 

CaO 

1 

4.130 

4  days 

+0.010 

5.419 

1.377 

0.686 

0.681 

0.139 

+0.912 

+0.557 

+0.344 

400  c.c.  milk;  23.56 

2 

. 

gm.  dried  meat. 

4  days 

-0.015 

5.419 

1.377 

0.686 

0.688 

0.217 

+0.567 

+0.472 

+0.350 

400  c.c.  milk;  23.56 

3 

gm.  dried  meat. 

4  days 

-0.005 

5.419 

1.377 

0.686 

0.677 

0.227 

+0.444 

+0.499 

+0.266 

400  c.c.  milk;  23.56 

4 

grm.  dried  meat. 

2  days 

+0.025 

5.419 

1.765 

1.212 

0.656 

0.403 

+0.784 

+0.7U 

+0.575 

Same  as  above  + 

5 

1  grm.  bone  dust. 

2  days 

-0.010 

5.419 

1.377 

0.686 

0.667 

0.361 

+0.514 

+0.350 

+0.087 

400  c.c.  milk;  23.56 
gm.  dried  meat. 

1 

3.600 

4  days 

+0.013 

4.781 

1.097 

0.524 

0.575 

0.114 

+0.688 

+0.408 

+0.176 

300  c.c.  milk;  23.56 

2 

gm.  dried  meat. 

4  days 

-0.005 

4.778 

1.184 

0.541 

0.638 

0.180 

+0.798 

+0.366 

+0.148 

300  c.c.  milk;  15.06 
gm.  dried  meat; 

3 

53.5  gm.  egg. 

4  days 

-0.013 

4.781 

1.097 

0.524 

0.611 

0.353 

+0.295 

+0.132 

+0.177 

300  c.c.  milk;  23.56 

4 

gm.  dried  meat. 

2  days 

+0.020 

4.781 

1.485 

1.050 

0.548 

0.717 

+0.115 

+0.220 

+0.001 

300  c.c.  milk;  23.56 
gm.   dried  meat; 

5 

1  gm.  bone  dust. 

2  days 

0.000 

4.781 

1.097 

0.524 

0.625 

0.379 

+0.160 

+0.094 

+0.288 

300  c.c.  milk;  23.56 
gm.  dried  meat. 

Age  of  dogs,  214-3  months. 

Since  the  intake  was  not  maintained  constant,  there  was  not  an 
entirely  satisfactory  basis  for  comparison  of  phosphorus  in  the  dif- 
ferent forms.  With  one  dog  the  bone  dust  caused  increased  reten- 
tion of  nitrogen,  calcium  and  phosphorus,  with  no  increase  in  urin- 
ary phosphorus,  but  naturally  a  decided  increase  in  feces  phospho- 
rus. With  the  other  dog  the  bone  dust  caused  diarrhoea,  and  the 
results  are  perhaps  affected  by  this  disorder.  Because  of  the  rapid 
decline  in  the  rate  of  phosphorus  storage  during  the  experiment 
with  the  second  dog,  it  is  impossible  to  say  just  what  was  the  effect 
of  the  egg  on  phosphorus  storage. 

Gilbert  and  Posternak  (1905)  compared  organic  and  inorganic 
phosphorus  compounds  in  balance  experiments  with  human  subjects. 
A  part  of  the  data  are  in  the  following  table. 


824 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


DAILY  NITROGEN  AND  PHOSPHORUS  BALANCES  FROM  A  NORMAL 

HUMAN  SUBJECT  WITH  ORGANIC  AND  INORGANIC  PHOSPHORUS 

COMPOUNDS  ADDED  TO  A  NORMAL  RATION 

Periods  of  Five  Days  Each — Grams 


Nitrogen 

Phosphorus  (P2O5) 

Ration 

Pood 

Urine 

Feces 

Balance 

Food 

Urine 

Feces 

Balance 

L    300  g-m.  meat;  450  g-m.  dried 

17.96 

17.72 

2.45 

—2.21 

2.43 

1.972 

0.675 

-0.217 

bread;  30  g-m.  butter;  100  gm. 

sugar;  1600  c.c.  tea. 

2.     Same  as  above  plus  mono-  and 

17.96 

16.62 

2.33 

-0.99 

3.19 

2.044 

1.337 

-0.191 

di-calcium  phosphates,  0.24  and 

0.52  gm.  P205- 

Fore-period,     constant  diet  for  5 

days. 
Same  as  above  plus  0.593  gm.  P2O5 

17.96 

15.85 

2.40 

-0.29 

2.43 

1.916 

0.643 

-0.129 

17.96 

16.96 

2.11 

—1.11 

3.02 

2.272 

0.853 

-0.102 

as  calcium  glycerophosphate. 

Same   as   above    without    added 

17.96 

16.24 

2.42 

-0.70 

2.43 

1.932 

0.683 

-0.185 

phosphorus. 

Same  as  above  plus  1  gm.  per  day 

17.96 

16.94 

2.20 

-1.18 

3.43 

2.100 

1.209 

+0.121 

P2O5  as  calcium  phytate. 

Age  of  subject,  3  years. 

In  this  experiment  the  phosphates  and  the  glycerophosphate 
were  not  well  retained,  though  they  did  serve  slightly  to  reduce  the 
phosphorus  loss.  Calcium  phytate,  however,  brought  about  de- 
cidedly improved  phosphorus  retention  and  a  positive  phosphorus 
balance.  The  amount  of  phosphorus  taken  in  this  form,  however, 
was  much  greater  than  in  the  other  cases,  so  that  we  do  not  have  an 
entirely  fair  basis  for  comparison.  In  consideration  of  the  char- 
acter of  the  basal  ration,  which  must  have  been  very  low  in  calcium, 
it  would  also  be  important  to  know  the  calcium  contents  of  the 
phosphatic  supplements. 

Gouin  and  Andouard  (1905-6),  in  calf -feeding  experiments, 
used  potassium  phosphate,  bone  phosphate,  sweetbreads,  thymus 
and  protylin  as  sources  of  phosphorus.  The  nature  of  the  data 
did  not  give  a  basis  for  a  close  estimate  of  the  values  of  these  com- 
pounds. The  authors  make  the  statement,  however,  that  they 
found  the  bone  phosphates  more  "digestible"  than  vegetable  phos- 
phates. In  this  connection,  we  would  mention  the  fact 
that  we  have  no  means  of  determining  the  digestibility  of  such  com- 
pounds. 

Tunnicliffe  (1906)  conducted  balance  experiments  with  two 
healthy  children,  aged,  respectively,  2  years,  and  2  years  and  10 
months,  comparing  organic  and  inorganic  phosphorus  compounds, 
and  their  effects  on  nitrogen  metabolism.  A  part  of  the  data  are 
as  follows: 


PHOSPHORUS  METABOLISM 


325 


AVERAGE  DAILY  NITROGEN  AND  PHOSPHORUS  BALANCES  WITH 

HEALTHY  CHILDREN  RECEIVING  ORGANIC  AND  INORGANIC 

PHOSPHORUS  WITH  THE  FOOD— Grams 


Subject 

Period 

Duration 

of  periods 

in  days 

N 

Food 

Urine 

Feces 

Balance 

P 

Food 

Urine 

Feces 

Balance 

Diet 

Boy,  2  years  old 

Fore-period 

2 

5.95 
2.21 
0.98 

+2.75 

0.69 
0.13 
0.18 

+0.38 

Mixed  normal  diet. 

Organic  phos- 
phorus period 

6 

8.80 
3.93 

0.78 
+4.08 

0.96 

0.27 

0.20 

+0.49 

Same  plus  20  gm.  sanatogen. 

Inorganic 

phosphorus 

period 

3 

5.76 
3.39 

0.88 
+J.48 

Same  as  first  plus  1  gm. 
Ca3  (P04>2. 

Girl,  2  yrs.  10  mos,  old 

Fore-period 

3 

6.53 

3.22 

0.75 

+2.56 

0.73 
0.31 
0.15 

+0.27 

Mixed  normal  diet. 

Girl,  2  yrs.  10  mos.  old 

Organic  phos- 
phorus period 

6 

9.14 
4;  70 
0.72 

+3.72 

0.99 
0.43 
0.12 

+0.44 

Same  plus  20  gm.  sanatogen. 

Girl,  2  yrs.  10  mos.  old 

Inorganic 

phosphorus 

period 

3 

5.80 

2.62 

0.76 

+2.42 

0.87 

0.28 

0.24 

+0.35 

Same  as  first  plus  1    gm. 
Ca3  (P04)2. 

From  these  data  Tunnicliffe  concluded  (1)  that  in  the  healthy- 
child  the  addition  of  an  organic  phosphorus  compound  to  the  diet  is 
followed  by  an  increase  in  the  amount  of  phosphorus  assimilated  by 
and  retained  in  the  body;  (2)  that  the  addition  of  an  organic  phos- 
phorus compound  to  the  diet  of  children  increases  the  amount  of  ni- 
trogen assimilated;  (3)  that  the  addition  of  Ca,(P04)2  to  the  food 
did  not  increase  the  amount  of  phosphorus  assimilated  or  retained 
by  the  child,  nor  did  this  compound  exert  any  favorable  influence 
upon  the  assimilation  of  the  nitrogen  of  the  food ;  and  (4)  that  the 
phosphorus  contained  in  the  sodium  glycerophosphate  of  casein 
(sanatogen)  is  practically  entirely  assimilated  by  the  body. 

We  would  suggest  that  the  periods  are  too  short  to  give  results 
of  great  value,  and  that,  since  the  intake  of  nitrogen  and  phosphorus 
was  greater  in  the  organic  phosphorus  period  than  in  the  inorganic 
phosphorus  period,  these  data  do  not  furnish  a  fair  basis  for  a  com- 
parison of  the  nutritive  values  of  these  compounds.  At  the  same 
time  it  seems  probable  that  the  phosphorus  of  sanatogen  is  more 
useful  than  the  same  amount  of  phosphorus  in  Ca3(P04)2. 


326 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


LeClerc  and  Cook  (1906)  conducted  a  series  of  nitrogen  and 
phosphorus  balance  experiments  with  three  rabbits  and  a  dog,  com- 
paring phytin  and  inorganic  phosphates,  in  twenty-six  five-day  peri- 
ods. The  phytin  was  prepared  from  wheat  bran.  This  compound 
and  a  mixture  of  disodium  hydrogen  phosphate  and  dihydrogen 
sodium  phosphate  were  added  to  normal  and  to  phosphorus-poor 
rations. 

From  the  considerable  weight  of  the  subjects  it  would  appear 
that,  if  not  mature,  they  were  at  least  beyond  the  period  of  most 
active  growth.  During  these  experiments  the  live  weights  were 
practically  constant ;  in  half  of  the  periods  there  was  loss  of  weight, 
and  in  none  of  them  was  there  marked  gain  in  weight.  These  con- 
ditions, together  with  the  brevity  of  the  periods,  were  unfavorable 
to  the  demonstration  of  such  differences  in  value  as  exist  between 
these  nutrients. 

The  authors  state  that  the  nitrogen  retention  was  generally 
lowered  by  the  addition  of  inorganic  phosphates,  when  fed  with  a 
normal  food ;  and  that  the  organic  phosphorus  compound  from 
wheat  bran  is  more  favorable  to  nitrogen  and  phosphorus  retention 
than  is  inorganic  phosphorus.      See  also  Cook  (1909),  p.  312. 

Egbert  Koch  (1906)  investigated  the  question  of  synthesis  of 
phosphorus  compounds  from  edestin  and  inorganic  salts  by  the  hu-! 
man  being,  by  means  of  a  feeding  experiment  on  a  man.  The  re- 
sults are  shown  in  the  following  table.  The  subject  was  in  a  normal 
state  of  nutrition  throughout  the  investigation. 

AVERAGE    DAILY   NITROGEN,    CALCIUM    AND    PHOSPHORUS 

METABOLISM  OF  A  MAN  ON  RATIONS  DIFFERING  IN 

ORGANIC    PHOSPHORUS    CONTENT 

Periods  of  Four  Days  Each — Grams 


Rations 


N 

Intake 

Urine 

Feces 

Balance 


P2O5 

Intake 

Urine 

Feces 

Balance 


CaO 

Intake 

Urine 

Feces 

Balance 


Weight 

Initial 

Final 

Difference 

Diff .  per  day 

Kg. 


100  gin.  oatmeal;  1500  c.c  milk; 
6  gm.  NaCl;  20  gm.  cane  sugar; 
100  gm.  butter:  500  c.c.  tea; 
300  c.c.  wine;  150  gm.  egg  white. 


100  gm.  oatmeal;  48.5  gm.  edestin; 
6  gm.  NaCl;  70  gm.  cane  sugar; 
180  gm.  butter;  500  c.c.  tea; 
300  c.c.  wine;  150  gm.  egg  white; 
13.36  gm.  Na2HP04;  4.12  gm.  CaCO.3; 
1.5gm.CaHP04. 


13.46 

10.31 

1.10 

+2.05 


4.43 

2.20 

1.90 

+0.34 


2.991 

0.137 

2.500 

+0.354 


63.9 

63.0 

-0.9 

-0.22 


12.97 

10.10 

0.97 

+1.65 


4.44 

2.98 

1.42 

+0.04 


2.997 
1.420 
3.038 
-0.183 


63.0 
63.7 
+0.7 
+0.18 


Calculated  from  author's  data. 


In  the  first  ration  the  phosphorus  was  present  to  a  considerable 
extent  as  phosphoprotein,  that  is,  as  casein.  In  the  second  ration 
the  phosphorus  was  present  principally  as  inorganic  phosphates. 


PHOSPHORUS  METABOLISM 


327 


The  intake  of  nitrogen,  phosphorus  and  calcium  was  maintained 
practically  constant,  though  the  nitrogen  intake  in  the  second  peri- 
od was  a  little  less  than  in  the  first. 

In  the  inorganic  phosphorus  period  the  nitrogen  storage  was 
less,  as  also  was  the  phosphorus  storage,  while  the  calcium  balance 
changed  from  +0.354  gm.  to  — 0.183  gm.  From  these  data  the 
author  concluded  that  "the  view  that  the  human  organism  cannot 
effect  a  synthesis  from  phosphorus-free  protein  and  inorganic  phos- 
phorus salts  receives  further  support  from  these  experiments." 

Hart,  McCollum  and  Fuller  (1909)  studied  the  role  of  inorganic 
phosphorus  in  the  nutrition  of  swine  by  feeding,  slaughter  and  bal- 
ance experiments.  The  daily  rations  in  their  first  experiment  were 
as  follows: 


DAILY  RATION  AND  AVERAGE  GAIN  IN  WEIGHT 

Lot  1 
Pounds 

Lot  2 
Pounds 

Lot  3 
Pounds 

Lot  4 
Pounds 

Lot  5 
Pounds 

1.24 
0.65 
0.26 

0.048 

1.20 
0.63 
0.25 

0.046 

0.077 

1.22 

0.64 
0.26 

0.047 

0.038 

1.26 
0.27 

0.67 

Ground  oats  0.67 

Sugar-salt  mixture  (200  gm.  sugar;   100  gm.  each 
NaCl,  MgCl2,  K2  SO4) 

Wheat  middlings 

Precipitated  calcium  phosphate  (Ca3  (P04)2  + 
CaHP04> 

0.67 
Oil  meal  0.22 

Grams 
1.12 

Grams 
6.57 

Grams 
3.84 

Grams 
5.40 

Grams 
5.45 

Pounds 
28.33 

Pounds 
52.6 

Pounds 
52.6 

Pounds 
59.0 

Pounds 
61.5 

There  were,  all  told,  16  pigs  in  this  experiment.  The  experi- 
ment covered  95  days,  after  which  one  animal  from  each  lot  was 
killed,  and  some  parts  were  analyzed  for  dry  matter,  calcium  and 
phosphorus.      The  calcium  and  phosphorus  data  are  as  follows : 


COMPOSITION  OF  PARTS  OF  PIGS- 

-Air-Dry  Basis — Percents 

Bone  ash 

Blood 

Leg  muscle 

Liver 

Brain 

P 

Ca 

P 

Ca 

P 

Ca 

P 

Ca 

P 

Ca 

Basal,  lotl 

Inorganic,  lot  2 . . . 
Whole  bran,  lot  4 . . 
Normal,  lot  5 

18.48 
18.26 
18  00 
18.20 

37.16 
36.91 
37.12 
37.23 

0.24 
0.31 
0.33 
0.28 

0.035 
0  031 
0.038 
0.026 

0.93 
0.81 
0.77 
0.78 

0.030 
0.029 
0.025 
0.041 

1.43 
1.34 
1.35 
1.27 

0.020 
0.030 
0.066 
0.030 

1.49 
1.57 
1.54 
1.43 

0.08 
0.10 
0.09 
0.09 

The  skeleton  of  one  animal  from  each  lot,  except  the  third,  was 
dissected  out  and  subjected  to  study  as  indicated  by  the  following 
data: 


328 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


DATA  ON  SKELETON  OF  ONE  ANIMAL  FROM  EACH  LOT  OF 
EXPERIMENT  I  (EXCEPT  LOT  3) 


Weight  of  skeleton,  gm 

Weight  of  animal,  lbs 

Breaking  strength  of  thigh  bone,  lbs.  per  sq.  mm 

Diam.  thigh  bone  at  centre,  mm 

Sp.  gr.  of  bones 

Ash  (thigh  bone) 


Basal 

Phosphate 

Whole  bran 

1193 

2371 

1288 

84 

123 

102 

0.63 

1.80 

1.84 

18.00 

23.90 

18.50 

0.977 

1.157 

1.100 

31  percent 

55  percent 

53  percent 

Standard 


1609 
138 
1.69 
22.00 
1.192 
46  percent 


Experiment  II  was  similar  to  the  above,  except  that  near  the 
end  of  the  program  one  animal  from  each  of  five  lots  was  put  into 
a  metabolism  crate,. and  subjected  to  a  balance  experiment  of  5-12 
days'  length.      The  whole  program  covered  123  days. 

In  this  experiment  the  rations  used  were  as  indicated  below : 

Lot  1 — Low  phosphorus  basal  ration. 
"  2 —  "  "  "  "       -f-Predprtated  calcium 

phosphate 
"3—"  "  "  "       +boneash 

"  4—  "  "  "  "       +floats  (rock  phosphate) 

"  5 — Basal  ration  with  whole  bran  substituted  for  washed  bran. 
"  6 — Normal  foods. 
The  summarized  balance  data  are  as  follows: 

AVERAGE  DAILY  PHOSPHORUS  BALANCES 


Phosphorus 
retention 


Basal 

Precipitated  phosphate 

Bone  ash 

Floats 

Whole  bran 


Aver,  gain 

Total 

Total 

Total 

Inorganic 

in  weight 

phosphorus 

phosphorus 

phosphorus 

phosphorus 

of  lot 

intake 

in  urine 

'in  feces 

in  feces 

Pounds 

Grams 

Grams 

Grams 

Grams 

32 

1.08. 

0.02 

0.52 

0.168 

42 

5.02 

0.378 

2.45 

35 

4.08 

0.281 

2.26 

43 

4.26 

0.253 

1.65 

58 

5.65 

0.666 

2.66 

2.47 

0.53 
2.22 
1.54 
2.35 
2.36 


One  animal  from  each  lot  in  this  experiment  was  killed,  and  be- 
low are  data  from  the  examination  of  the  skeletons. 

DATA  ON  THE  SKELETON  OF  ONE  ANIMAL  FROM  EACH  LOT  OF 
EXPERIMENT  II  (EXCEPT  LOT  6) 


Weight  of  skeleton,  gm , 

Weight  of  animal  at  slaughter,  lbs 

Breaking  strength  of  thigh  bone,  lbs.  per  sq.  mm 

Diam.  thigh  bone  at  centre,  mm 

Sp.  gr.  bone 

Ash)  percent  (thigh  bone) 


870 

77 
0.87 
16.0 
0.984 

33 


Precipitated 
phosphate 


950 
87 

1.70 
16.0 

1.15 
46 


950 
58 

1.77 
15.5 

1.12 
53 


Floats 


1495 

82 

1.65 

20.00 

1.19 

57 


Whole 
bran 


850 
87 

1.86 
17.00 

1.14 
54 


PHOSPHORUS  METABOLISM  829 

The  authors'  conclusions  are  as  follows : 

"1.  On  the  ration  extremely  low  in  phosphorus,  pigs  made  as 
large  gains  up  to  75  or  100  pounds  when  starting  at  weights  of  from 
40  to  50  pounds  as  animals  receiving  an  abundance  of  this  element. 
After  reaching  this  point  loss  of  weight  began,  followed  by  collapse. 

"2.  When  such  low  phosphorus  rations  as  induced  the  above 
symptoms  were  supplemented  with  calcium  phosphates,  no  un- 
toward results  appeared.  Animals  fed  a  low  phosphorus  ration, 
supplemented  with  inorganic  phosphates,  made  as  vigorous  a  devel- 
opment as  others  receiving  their  phosphorus  supply  wholly  in  or- . 
ganic  form. 

"3.  Precipitated  calcium  phosphates,  a  mixture  of  di-  and  tri- 
calcium  phosphates,  gave  no  better  results  than  did  floats,  a  crude 
tri-calcium  phosphate. 

"4.  Phytin  as  the  supply  of  phosphorus  gave  no  better  re- 
sults than  the  inorganic  phosphates. 

"5.  A  young  animal  of  40  pounds  weight  receiving  inorganic 
phosphates,  together  with  other  salts  as  supplementary  to  a  ration 
very  low  in  mineral  constituents,  grew  to  be  an  animal  of  280  pounds 
weight,  bore  a  litter  of  fairly  vigorous  pigs,  which  on  the  same  ra- 
tion completed  the  cycle  back  to  80  pounds,  while  animals  on  the 
same  ration  less  the  inorganic  phosphates  collapsed  in  three  months, 
with  loss  of  weight  accompanied  by  a  loss  of  the  use  of  their  limbs. 

"6.  Determinations  of  calcium  and  phosphorus  on  the  prin- 
cipar  organs  and  tissues  of  the  animals  on  the  low  phosphorus  ra- 
tion showed  that  they  maintained  the  proportion  of  these  elements 
constant  and  comparable  to  that  of  normally  fed  pigs. 

"7.  The  percentage  of  ash  in  the  skeleton  of  pigs  on  the  de- 
pleted phosphorus  ration  was  reduced  to  nearly  one-half  that  of  pigs 
receiving  a  normal  ration,  or  a  phosphorus-poor  ration  supplemented 
by  an  inorganic  phosphate. 

"8.  The  marked  reduction  in  the  quantity  of  ash  of  the  bones 
of  the  animal  receiving  an  insufficient  supply  of  calcium  phosphates, 
together  with  the  ability  of  the  animal  to  build  up  a  skeleton  very 
rich  in  calcium  phosphate  when  an  abundance  of  the  latter  is  sup- 
plied in  inorganic  forms,  strongly  points  to  the  possession  of  a  syn- 
thetic power  by  the  animal  which  enables  it  to  convert  inorganic 
forms  of  phosphorus  into  the  organic  forms  demanded  by  its  body. 

"9.  When  animals  were  starving  for  phosphorus,  they  drew 
this  element  from  the  skeleton,  but  removed  calcium  and  phospho- 
rus in  the  proportions  found  in  tri-calcium  phosphate. 


330  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

"10.  The  daily  phosphorus  supply  for  a  50-pound  growing"  pig* 
should  be  at  least  3  gm.  A  supply  of  4  or  5  gm.  is  probably  a  safer 
quantity. 

"11.  The  data  furnish  no  positive  evidence  of  the  synthesis  of 
nucleo-proteids  or  other  phosphorus-bearing  complexes  from  inor- 
ganic phosphates  in  the  animal  body." 

On  these  conclusions  we  would  offer  the  following  comments: 

The  basal  ration  used  in  these  experiments  was  exceedingly  low 
in  calcium.  To  have  furnished  a  wholly  satisfactory  basis  for  the 
comparison  of  the  phosphorus  compounds  studied  there  should  have 
been  no  question  as  to  calcium  deficiency  limiting  the  usefulness  of 
any  of  these  preparations.  In  this  light,  the  phosphorus  of  floats 
had  the  greatest  advantage,  since  it  was  associated  with  an  excess 
of  calcium;  the  phosphorus  of  precipitated  phosphate  came  next  in 
order  of  relation  of  calcium  to  phosphorus,  while  the  wheat  bran 
phosphorus  was  associated  with  an  exceedingly  small  proportion  of 
calcium. 

The  authors'  statement  that  the  calcium  and  phosphorus  con- 
tent of  the  principal  organs  of  the  body  was  maintained  constant  is 
not  sustained  by  their  analytical  data.  True,  there  is  a  marked 
tendency  toward  the  maintenance  of  constancy  of  composition,  but, 
at  the  same  time,  there  are,  in  the  case  of  each  of  the  five  important 
parts  analyzed,  variations  in  the  content  of  calcium  and  phosphorus, 
which,  reckoned  on  the  basis  of  the  total  amount  of  the  constituent 
present,  constitute  marked  deviations  from  the  normal. 

The  authors'  conclusion  No.  8,  citing  the  utilization  of  inor- 
ganic calcium  phosphate  in  the  building  up  of  the  skeleton  as  an  evi- 
dence of  "the  possession  of  a  synthetic  power  by  the  animal  which 
enables  it  to  convert  inorganic  forms  of  phosphorus  into  the  organic 
forms  demanded  by  its  body,"  seems  not  to  be  warranted.  Con- 
clusion No.  11,  which  claims  "no  positive  evidence  of  synthesis  of 
nucleo-proteids  or  other  phosphorus-bearing  complexes  from  inor- 
ganic phosphates  in  the  animal  body,"  is  more  to  the  point. 

These  experiments  seem  not  well  planned  to  bring  out  differ- 
ences in  usefulness  of  organic  and  inorganic  phosphorus.  They 
do  make  emphatic  one  point,  however;  that  the  amount  of  organic 
phosphorus  absolutely  necessary  to  the  life  of  the  pig  is,  at  the 
most,  not  a  large  part  of  the  total  phosphorus  requirement. 

Holsti  (1910)  attempted  to  settle  the  question  of  organic  phos- 
phorus synthesis  by  balance  experiments  on  himself;  but,  as  the 
rations  differed  in  total  phosphorus  content  by  as  much  as  the 
amounts  added  to  some  of  them  as  inorganic  phosphorus,  there  was 


PHOSPHORUS  METABOLISM  331 

in  reality  no  basis  whatever,  for  judgment  as  to  the  matter  of  syn- 
thesis of  organic  phosphorus  compounds.  This  much  was  demon- 
strated, however,  that  if  the  total  phosphorus  of  the  ration  is  ad- 
equate, a  very  small  intake  of  organic  phosphorus  (about  a  third  of 
a  gram  sufficed  in  one  case)  will  permit  of  phosphorus  retention, 
even  coincident  with  nitrogen  loss.  Holsti's  experimental  periods, 
3-6  days  in  length,  were  too  short  really  to  settle  anything. 

Under  the  title  "Nuclein  Synthesis  in  the  Animal  Body," 
McCollum  (1909)  published  results  of  feeding  experiments  with  rats 
on  a  basal  ration  composed  of  phosphorus-free  foods,  to  which  were 
added  phosphorus-containing  substances  to  be  compared.  Among 
the  foods  used  were  edestin,  zein,  corn  starch,  wheat  starch,  butter 
fat,  bacon  fat,  milk  sugar,  glucose,  cane  sugar,  and  cholesterin.  Many 
flavoring  extracts  were  used  in  an  effort  to  make  the  food  palatable. 
Milk  ash,  calcium  phosphate,  sodium  chloride  and  ferric  chloride 
were  also  used  in  the  basal  ration,  and  casein,  and  hydrolyzed  beef 
and  liver  were  added  to  this  ration  in  some  periods. 

McCollum  concludes  that,  other  things  being  satisfactory,  all 
the  phosphorus  needed  by  an  animal  for  skeleton,  nuclein,  or  phos- 
phatid  formation,  can  be  drawn  from  inorganic  phosphates;  also 
that  the  animal  has  the  power  to  synthesize  the  purin  bases  neces- 
sary for  its  nuclein  formation  from  some  complexes  contained  in  the 
protein  molecule,  and  does  not  necessarily  use  purin  bases  of  exogen- 
ous origin  for  this  purpose.  McCollum  also  placed  much  emphasis 
on  the  idea  of  palatability  as  of  vital  importance  in  nutrition. 

Rats  1,  2  and  3  were  fed  a  normal  ration,  and  gained  in  weight 
one  gram  or  more  per  day.  Rats  4,  5  and  6  were  fed  on  the  organ- 
ic-phosphorus-free ration  and  lost  weight  rapidly.  Rats  7,  8  and  9 
received  the  same  ration  as  the  above,  but,  in  addition,  protein-free 
hydrolyzed  beef  and  liver.  Rats  8  and  9  lost  27  and  28  grams,  re- 
spectively, in  106  days.  Rat  7  gained  23  grams  in  53  days.  It  was 
put  into  the  experiment  on  June  12,  weighing  153  grams.  By  June 
23  it  weighed  168  grams.  On  July  28,  thirty-five  days  later,  it 
weighed  170  grams,  though  between  these  dates  it  had  weighed  as 
much  as  180  grams.  On  Aug.  4  it  was  removed  from  the  experi- 
ment at  a  weight  of  176  grams.  Thus  far,  then,  five  out  of  six  rats 
had  failed  to  maintain  their  weight;  the  sixth  had  made  a  gain  in 
weight. 

Rats  18,  19  and  20  were  younger  than  those  used  thus  far. 
They  were  fed  on  the  same  ration  as  rats  4,  5  and  6,  that  is,  with- 
out organic  phosphorus.  They  all  gained  in  weight.  At  the  be- 
ginning of  the  test  they  weighed  37,  35  and  46  grams.      In  127,  56 


332 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


and  56  days,  respectively,  they  gained  39,  6  and  14  grams,  respect- 
ively, in  weight.  The  one  rat  which  more  than  doubled  its  weight 
was  analyzed,  along  with  certain  others,  for  dry  substance  in  the 
body,  fat  in  the  dry  matter,  and  weight  and  ash  of  skeleton.  (See 
table  below.) 

Two  rats,  Nos.  15  and  16,  were  fed  on  the  same  ration  plus 
casein,  and  gained  in  weight  without  difficulty. 

COMPOSITION  OF  RATS  USED  IN  EXPERIMENTS  WITH  VARIOUS 

RATIONS 


Fat-  and 

Dry  tis- 

Ether 

Ash  of 

Skeleton 

water-free 

Num- 

Weight 

Skeleton 

sues  less 

extract 

skeleton 

tissues 

Ration 

ber  of 
rat 

Grams 

Grams 

skeleton 
Grams 

Grams 

Grams 

Percent  of 
live  weight 

Percent  of 
live  weight 

1 

2 
10 

147 

157 

34 

6.67 
6.50 
1.33 

38.0 

45.0 

9.5 

8.89 

10.80 

3.25 

3.79 

3.85 
0.68 

4.54 
4.14 
3.91 

19.8 

21.79 

18.39 

Inorganic  phosphorus: 

later  phosphorus-free 

7 

102 

7.14 

25.5 

3.42 

4.49 

7.00 

21.64 

Inorganic  phosphorus 

4 

135 

9.00 

34.8 

7.00 

4.28 

6.66 

20.59 

•  <                   << 

5 

96 

5.78 

22.0 

4.01 

3.02 

6.02 

18.94 

•  •                   <  • 

6 

88 

7.50 

20.5 

3.63 

3.18 

8.52 

19.17 

<  •                   • « 

8 

103 

6.03 

21.5 

2.67 

3.52 

5.85 

18.28 

<>                  i< 

9 

78 

4.58 

14.5 

0.86 

2.77 

5.79 

17.49 

18 

76 

4.07 

17.5 

3.40 

2.14 

5.36 

18.55 

By  comparing  the  figures  from  No.  18,  the  one  rat  which 
doubled  its  weight,  with  those  from  Nos.  1  and  2,  which  were  fed  on 
normal  foods,  one  may  satisfy  himself  by  computation  that  the  gain 
in  weight  of  this  one  rat  could  not  be  made  up  entirely  of  fat,  water, 
intestinal  content  and  skeleton.  The  extent  to  which  the  gain  in 
weight  was  dependent  on  translocation  of  constituents  was  not  de- 
termined. There  were  no  determinations  of  moisture  in  the  tis- 
sues, nor  of  nuclein  or  lecithin  phosphorus,  and  there  were  no 
weights  of  foods  taken.  The  author  states  in  a  letter  that  there 
was  opportunity  for  the  rats  to  eat  their  own  feces,  a  fact  which  we 
have  found  enters  in  most  important  ways  into  the  determination  of 
results  in  such  work,  apparently  first  through  allowing  of  repeated 
use  of  the  lecithin  of  the  bile  residues,  and  second  through  allowing 
the  animal  to  avail  itself  of  results  of  the  synthetic  capacities  of 
intestinal  bacteria. 

The  nature  of  the  analytical  data  hardly  warrants  the  author's 
assumption  that  constancy  of  composition  of  the  tissues  was  main- 
tained, and  his  reference  to  the  work  of  Hart,  McCollum  and  Fuller 
with  swine,  as  sustaining  the  assumption,  is  not  convincing. 

One  rat  was  so  confined  that  the  excreta  could  be  collected, 
and  a  ration  was  given  it  of  phosphorus-free  foods.  The  amounts 
eaten  were  not  determined.      As  an  average  of  8  days'  excretion, 


PHOSPHORUS  METABOLISM  333 

terminating  four  days  before  the  death  of  the  rat,  the  author  sub- 
mits the  figure  0.0063  gm.  phosphorus  as  the  daily  elimination  of 
this  rat,  which  weighed  180  gm.  at  its  maximum,  and  102  gm.  at  its 
death.  With  this  figure  as  a  basis,  the  author  computes  that  dur- 
ing the  course  of  his  experiments  certain  rats  must  have  entirely 
changed  the  phosphorus  content  of  the  body,  the  skeleton  excepted. 
He  assumes  that  the  rats  did  not  draw  phosphorus  from  the  bones 
from  the  fact  that  the  skeletons  increased  in  weight,  while  the  ani- 
mals lost  in  weight;  but  there  was  no  evidence  presented  to  show 
that  there  was  not  withdrawal  of  organic  phosphorus  from  the  skel- 
eton coincident  with  the  deposit  of  inorganic  phosphates. 

The  general  failure  of  the  older  rats  even  to  maintain  live 
weight  on  the  organic-phosphorus-free  diet,  taken  together  with  the 
ability  of  the  younger  rats  to  gain  in  weight  on  this  ration,  implies 
either  that  the  older  animals!  did  not  eat  food  enough,  or  else  that 
the  ability  of  the  rat  to  gain  in  weight  on  rations  which  are  free 
from  organic  phosphorus  compounds,  if  possessed  at  an  early  age, 
becomes  less  marked  with  advance  in  age. 

Gregersen  (1911)  conducted  an  extensive  series  of  balance  ex- 
periments with  rats,  the  object  being  to  study  phosphorus  metab- 
olism, especially  the  possibility  of  the  synthesis  of  organic  phos- 
phorus compounds  from  phosphorus-free  edestin  and  inorganic 
phosphates.  The  work  includes  in  all  48  experiments  involving  170 
balances.  The  method  of  handling  the  rats  was  that  of  Henriques 
and  Hansen  (Zeitschr.  physiol.  Chem.,  1905,  XLIII,  418).  The  data 
reported  include  analyses  of  food,  urine  and  feces,  but  no  analyses 
of  the  rats  themselves.  The  author's  tables  have  been  transcribed 
without  material  change  but  for  the  omission  of  the  separate  urine 
and  feces  data.     His  last  table  we  have  omitted. 

Gregersen  concludes  that  the  organism  is  able  to  synthesize 
organic  phosphorus  compounds  from  phosphorus-free  organic  ma- 
terials and  phosphates ;  in  feeding  on  a  nitrogen-free  ration  the  or- 
ganism is  not  affected  by  the  presence  or  absence  of  phosphates; 
when  the  organism  is  held  in  nitrogenous  equilibrium  on  a  phospho- 
rus-free albumin-containing  ration,  the  excretion  of  phosphorus 
falls  off  quite  considerably,  under  some  circumstances  becoming  as 
low  as  1/50 — 1/60  the  amount  of  the  coincident  nitrogen  excretion; 
on  a  phosphorus-free,  albumin-containing  ration,  which  contains  cal- 
cium and  magnesium  salts,  the  rats  excrete  only  a  minimum  amount 
of  phosphorus  through  the  urine,  less  than  one-tenth  of  that  quan- 
tity which  is  at  the  same  time  excreted  in  the  feces ;  on  a  phospho- 


334  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

rus-free  ration  containing  albumin  but  no  calcium  or  magnesium 
salts  more  phosphorus  is  usually  excreted  through  the  urine  than 
through  the  feces,  likewise  on  a  phosphorus-free  albumin-free  diet 
which  contains  calcium  and  magnesium.  It  is  not  claimed  that 
phosphorus-free  nitrogenous  compounds  and  phosphates  are  of 
equal  value  with  phosphorized  nitrogenous  compounds. 

From  a  superficial  inspection  of  the  data  one  is  inclined  to  agree 
with  Gregersen's  conclusion  as  to  the  synthesis  of  organic  phospho- 
rus compounds,  and  the  facts  may  be  as  concluded  by  Gregersen, 
but,  considering  the  mass  of  conflicting  testimony  which  has  been 
reported  on  this  subject,  we  must  weigh  the  evidence  with  care.  In 
certain  respects  the  proof  of  organic  phosphorus  synthesis  appears 
incomplete  or  imperfect.  There  were  a  great  number  of  experi- 
ments ;  but  the  periods  were  very  short.  The  data  would  be  much 
more  satisfactory  if  they  included  determinations  of  the  amounts  of 
organic  and  inorganic  phosphorus  in  the  bodies  of  the  experimental 
animals  and  in  controls.  Our  knowledge  of  the  capabilities  of  the 
animal  to  transfer  and  to  transform  phosphorus  from  one  tissue  or 
compound  to  another,  under  the  stress  of  necessity,  is  too  limited 
to  warrant  unreserved  statements  as  to  synthesis  of  individual  com- 
pounds when  such  deductions  rest  upon  only  the  storage  of  the  nu- 
trient and  gain  in  live  weight  during  periods  of  a  very  few  days.  Re- 
garding certain  points  one  is  left  in  doubt  since  the  percentage  com- 
position of  the  salt  mixtures  was  not  stated. 

Turning  to  Table  I,  p.  335,  these  rats  were  taken  from  a  bread 
diet,  and,  without  an  intermediate  period  on  the  experimental  ration 
to  free  them  from  the  effects  of  the  ration  of  white  bread,  but  after 
a  fast  from  one  afternoon  until  9 :00  o'clock  next  morning,  were  put 
into  the  balance  experiments,  in  which  an  abundance  of  protein  in 
the  form  of  edestin,  and  of  phosphorus  in  the  shape  of  phosphates, 
were  provided.  Now  bread  is  deficient  in  protein  and  in  phospho- 
rus, and  extremely  low  in  calcium,  so  low  that  it  could  hardly  have 
failed,  with  these  growing  rats,  to  have  caused  a  loss  of  calcium, 
and  therefore  probably  also  of  phosphorus,  from  the  bones,  if  indeed 
the  rats  received  nothing  but  bread.  The  abrupt  change  to  the  ex- 
perimental ration  found  the  rats  much  in  need  of  the  protein,  cal- 
cium and  phosphorus  which  this  ration  provided,  and  there  was  con- 
sequently, for  a  short  time,  a  marked  storage  of  these  constituents. 
This  apparent  prosperity,  however,  was  not  sustained. 


PHOSPHORUS  METABOLISM 


335 


TABLE  I.       METABOLISM  EXPERIMENTS  ON  GROWING  RATS  ON  A 
DIET  CONTAINING  NO  ORGANIC  PHOSPHORUS— Daily  Balances 


No.  of  investiga- 
tion and  weight  of 
subject  in  grams 

Length  of 

period  in 

days 

Food 
per  day 

Grams 

Food  N 
Milligrams 

Food  P 
Milligrams 

N  balance 
Milligrams 

P  balance 
Milligrams 

N:P 
retained 

1                          26 
1                          28 

5 
5 

2.76 
2.04 

71 
52 

9.7 
7.2 

23 
6 

3.5 
1.4 

6.6 

4.3 

2                          36 
2                          38 
2                        38 

5 
5 
5 

3.59 
2.58 
2.58 

92 
66 
66 

12.6 
9.1 
9.1 

32 
10 

4 

4.8 
2.3 
2.1 

6.7 
4.3 
1.9 

3                         49 
3                         54 
3                        55 

4 
4 
5 

5.20 
4.70 
3.71 

134 
121 
95 

18.3 
16.5 
13.1 

42 

17 

6 

7.1 
2.6 
2.2 

5.9 
6.5 
2.7 

4                        43 
4                        48 
4                        49 
4                         50 
4                        45 

4 
4 
5 
5 
5 

4.95 
3.56 
2.85 
2.02 
2.48 

127 
91 
73 
52 
64 

17.4 

12.5 

10.0 

7.1 

8.7 

42 

20 

10 

—24 

—11 

7.3 
2.5 
1.5 
—1.4 
0.0 

5.8 
8.0 
6.7 

5                         44 
5                         50 
5                        51 

5 
5 
5 

4.12 
3.51 
2.92 

106 
90 

75 

14.5 

12.4 
10.3 

31 

12 

—  4 

6.1 

1.8 

-0.5 

5.1 
6.7 

6                         63 
6                        71 
6                         66 
6                         65 

6 
6 
6 
6 

4.57 
3.21 
3.19 
2.91 

117 

83 

82      • 
75 

16.1 
11.3 
11.2 
10.2 

35 
3 
1 
3 

3.9 
1.3 
1.2 
1.9 

9.0 
2.3 
0.8 
1.6 

7                         60 
7                        68 
7                       62 
7                       62 

5 

6 

6 

'   6 

4.96 
2.92 
3.47 

2.85 

128 
75 
89 
73 

17.4 
10.3 
12.2 
10.0 

30 

—  7 

2 

-10 

4.0 
0.3 
0.9 
0.5 

7.5 
2.2 

8                         58 
8                        63 
8                         68 

6 

7 
7 

5.38 
4.50 
3.83 

138 
116 
98 

18.9 
15.8 
13.5 

37 

26 

6 

4.3 
4.6 
3.3 

8.6 

5.7 

1.8 

The  ration  was  composed  of  edestin  15  percent,  sugar  30  percent,  fat  42  percent, 
cellulose  5  percent,  sodium  phosphate  4=  percent,  and  4  percent  of  a  salt  mixture  containing 
sodium,  potassium,  and  calcium  chlorides,  sodium  bicarbonate,  magnesium  oxide  and  iron 
sulphate. 

In  these  8  series  of  balances  there  was  in  each  case  a  progres- 
sive decline  in  the  food  consumed,  and  in  the  nitrogen  and  phospho- 
rus storage.  These  8  experiments  were  10-24  days  in  length.  Three 
out  of  the  8  showed  minus  nitrogen  balances,  one  in  the  period  com- 
posed of  the  13th-18th  days,  another  during  the  10th-15th  days,  and 
the  third  during  the  5th-llth  days.  Two  of  these  minus  nitrogen 
balances  were  accompanied  by  negative  phosphorus  balances,  and 
in  the  third  the  positive  balance  was  but  0.0003  gm.  In  6  cases  out 
of  the  8  there  was  a  progressive  decrease  in  the  proportion  of  nitro- 
gen to  phosphorus  stored,  at  the  same  time  that  the  storage  of  both 
elements  also  declined,  as  above  noted. 

The  daily  nitrogen  storage  in  the  first  period  of  4-6  days  was 
23-42  mg. ;  during  the  second  period  the  storage  was  from  — 7  to 
+0.26  mg.,  and  in  the  third  period  from  — 4  to  +10  mg.  The  phos- 
phorus storage  in  the  first  period  was  from  3.5  to  7.3  mg.,  in  the  sec- 
ond, 0.3  to  4.6  mg.,  and  in  the  third  from  — 0.5  to  -f-3.3  mg. 


336  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

These  facts  are  probably  due  to  the  falling-  off  in  food  consump- 
tion, which,  of  course,  would  leave  the  rat  with  a  smaller  amount  of 
protein,  in  excess  of  the  maintenance  requirement,  available  for 
storage,  and  also  a  smaller  proportion  of  the  total  protein.  The 
above-mentioned  decrease  in  food  consumption,  if  accompanied  by  a 
considerable  storage  of  phosphorus  in  the  bones  might  then  account 
for  the  decreased  proportion  of  nitrogen  to  phosphorus  which  was 
stored.  In  this  connection  we  would  observe  that  the  palatability  of 
a  ration  is  very  commonly  a  reflection  of  the  usefulness  of  the  food 
to  the  animal. 

The  general  results  of  this  first  series  of  balance  experiments 
suggest  great  difficulty  in  the  maintenance  of  normal  conditions  of 
growth.  This  test  is  the  most  thoroughgoing  of  any  in  the  whole 
investigation,  in  that  the  periods  were  much  longer  than  in  subse- 
quent series ;  also  the  rats  were  young,  growing  animals,  and  there- 
fore had  less  reserve  material  within  the  body  with  which  to  bridge 
over  temporary  nutritional  deficiencies. 

In  Table  II,  p.  337,  we  have' the  second  series  of  experiments. 
The  periods  were  of  three  to  five  days  duration.  Here  we  have  grown 
rats,  fed  with  and  without  protein,  and  without  organic  phosphorus 
compounds.  The  nitrogen-free  ration  was  composed  of  sugar,  lard, 
cellulose  and  salts,  including  sodium  phosphate.  The  ration  contain- 
ing protein  was  composed  like  the  above,  except  for  the  substitution 
of  edestin  for  15  percent  of  sugar.  The  albumin-free  ration  was 
the  same  as  the  nitrogen-free  ration,  except  for  the  addition  of  3 
percent  of  Liebig's  extract  of  beef. 

In  Experiments  9-22  there  are  18  changes,  either  from  a  nitro- 
gen-free ration,  or  from  one  in  which  the  only  nitrogen  present  was 
in  meat  extractives,  to  a  ration  where  the  sole  albuminous  nitrogen 
present  was  in  edestin,  or  changes  in  the  reverse  direction.  Sodium 
phosphate  was  present  in  all  rations  whether  containing  protein  or 
without  protein.  In  each  of  these  18  changes  of  food  the  nitrogen 
and  phosphorus  balances  changed  consistently  from  -f-  to  — ,  or 
from  —  to  +>  according  to  whether  or  not  edestin  was  present;  that 
is,  there  was  in  every  case  a  storage  of  both  nitrogen  and  phos- 
phorus in  the  presence  of  edestin,  and  in  no  case  a  storage  of  either 
element  in  the  absence  of  edestin,  even  though  the  phosphorus  was 
present  in  greater  amount  than  when  edestin  was  present. 

The  loss  of  phosphorus  during  albumin-free  periods  was  prob- 
ably due  to  the  katabolism  of  nitrogenous  tissues  which  contained 
phosphorus.  Then  with  the  introduction  of  edestin  into  the  ration, 
the  phosphorus-containing  proteins  were  protected,  and  there  oc- 
curred both  nitrogen  and  phosphorus  retention.  One  can  only  spec- 
ulate as  to  the  form  in  which  the  phosphorus  was  retained.      The 


PHOSPHORUS  METABOLISM 


337 


average  daily  loss  of  P  on  the  nitrogen-free  diet  during  the  whole 
series  was  7.0  mg.  The  daily  retention  on  the  edestin  ration  aver- 
aged 2.3  mg.  There  was  not  conspicuous  uniformity  in  the  ratio  of 
nitrogen  to  phosphorus  either  in  the  loss  or  the  retention  of  these 
elements. 

TABLE  II.      METABOLISM  EXPERIMENTS  ON  MATURE  RATS  WITH 

AND  WITHOUT  PROTEIN  AND  WITHOUT  ORGANIC 

PHOSPHORUS— Daily  Balances 


1   No.  of  investiga- 
tion, and  weight  of 
subject  in  grams 

Length 

of 
period 
in  days 

Food 
per  day 

Grams 

Food  N 

Milli- 
grams 

Food  P 

Milli- 
grams 

N 
balance 

Milli- 
grams 

P 

balance 

Milli- 
grams 

N:P 
retained 
or  lost 

Ration 

9                      101 
9                    107 

3 
3 

7.53 
7.53 

0 
197 

26.4 
26.6 

-63 
61 

-4.7 
2.9 

13.4 
21.0 

N-free 
Edestin 

10                     £39 
10                     224 
10                     228 

3 
3 
3 

10.00 
10.00 
8.50 

0 

261 
0 

35.0 
35.3 
29.8 

—137 
14 

—107 

-12.5 
0.9 

-9.0 

11.0 
15.6 
11.9 

N-free 

Edestin 

N-free 

11                     152 
11                     146 

3 
3 

10.00 
7.80 

0 

204 

35.0 

27.5 

—107 
33 

-7.3 

2.0 

14.7 

16.5 

N-free 
Edestin 

12                     195 
12                      188 
12                      196 

3 
3 
3 

9.00 
9.00 
9.00 

0 

235 
0 

31.5 
31.8 
31.5 

-81 
51 

-77 

-6.1 

4.6 

-4.4 

13.3 
11.1 
17.5 

N-free 

Edestin 

N-free 

13                      125 
13                      116 

3 
4 

6.50 
5.69 

0 
149 

22.8 
20.1 

-59 
23 

-6.6 
2.0 

8.9 
11.5 

N-free 
Edestin 

14                      138 
14 1                    131 
14                      137 
14                     vl36 

3 
3 
3 
3 

6.00 
6.00 
7.00 
5.67 

0 
157 

0 
148 

21.0 
21.2 
24.5 
20.0 

-57 
17 

-57 
26 

-4.9 
0.8 

-2.1 
1.3 

11.6 

21.4 
27.1 
20.0 

N-free 
Edestin 
N-free 
Edestin 

15                      160 
15                      157 

3 

4 

8.00 
6.14 

0 
160 

28.0 
21.7 

—63- 
27 

-6.0 

2.4 

10.5 
.    11.3 

N-free 
Edestin 

16                      58 
16                      55 

3 

4 

3.50 
3.50 

0 
91 

11.5 
11.6 

-44 
18 

-3.6 
2.2 

12.2 
8.2 

N-free 
Edestin 

17                      58 
17                      55 

3 
4 

3.55 
.3.18 

0 
83 

11.7 

10.5 

-45 
13 

-4.7 
1.8 

9.6 

7.2 

N-free 
Edestin 

18                      72 
18                   '   72 

3 
4 

5.00 
2.93 

0 
76 

16.5 
9.7 

-37 
1 

-2.0 
0.3 

18.5 
3.3 

N-free 
Edestin 

19                     290 
19                     283 
19                     288 
19                      307 

2 
1 
3 
1 

12.00 
12.00 
12.00 
12.00 

40 
304 
304 
304 

49.7 
39.3 
39.3 
39.3 

-171 

f. 

-16.2 
5.1 

10.6 
8.3 

Albumin-free 
Edestin 

20                      175 
20                      169 
20                      172 
20                      174 

2 
1 
2 
1 

8.00 
8.00 
8.00 
8.00 

26 

202 

'  202 

202 

33.1 
26.2 
26.2 
26.2 

—107 

-8.9 
0.7 

12.0 
26.4 

Albumin-free 
Edestin 

21                      320 
21                      317 
21                      322 
21                     335 

2 
1 
3 
1 

12.00 
12.00 
12.00 
12.00 

40 
304 
304 
304 

49.7 
39.3 
39.3 
39.3 

—133 

-12.0 
3.5 

11.1 
14.4 

Albumin-free 

Edestin 

22                     247 
22                    244 
22                    251 
22                    262 

2 
1 

2 
2 

11.00 
11.00 
11.00 
11.00 

36 

278 
278 
278 

45.5 
36.0 
36.0 
36.0 

-108 
J-    49 

-8.0 
4.5 

13.5 
10.9 

Albumin-free 
Edestin 

The  nitrogen- free  ration  was  composed  as  follows:  Sugar  45  percent,  lard  42  percent, 
cellulose  5  percent,  and  salts,  including  sodium  phosphate,  8  percent. 

The  edestin  ration  was  composed  as  above  except  that  15  percent  of  sugar  was  replaced 
by  edestin. 

The  albumin-free  ration  was  composed  like  the  above  nitrogen-free  ration  except  for 
the  addition  of  3  percent  of  Liebig's  beef  extract. 


888 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


The  experiments  in  Table  III  (below)  compare  powdered  beef 
and  sodium  phosphate  with  edestin  and  sodium  phosphate.  In  this 
series  the  edestin  period  in  every  case  follows  immediately  after  a 
meat  period,  without  intermediate  feeding,  certain  residua  from  the 
meat  feeding-  doubtless  holding  over  into  the  edestin  periods  in  such 
way  as  to  mix  results,  and  to  favor  the  utilization  of  the  edestin  ra- 
tions. This  advantage  was  increased  by  the  fact  that  the  prelim- 
inary meat  period  was  10-14  days  long,  the  balances  covering,  on  an 
average,  the  last  8.3  days  of  this  period,  while  the  following  edestin 
periods  were,  on  an  average,  only  3.8  days  long.  In  spite  of  these 
manifest  advantages  to  the  edestin  rations,  the  meat  rations  made 
a  slightly  better  showing  in  the  phosphorus  balances. 

TABLE  III.      METABOLISM  EXPERIMENTS  WITH  GROWN  RATS 
COMPARING  POWDERED  BEEF  WITH  EDESTIN— Daily  Balances 


No.   of 

tion,  an 
subject 

investiga- 
d  weight  of 
in    grams 

Length 

of 
period 

Food 
per  day 

Food  N 
Milli- 

FoodP 
Milli- 

N 
balance 

Milli- 

P 

balance 

Milli- 

N:P 

retained 

Ration 

in  days 

Grams 

grams 

grams 

grams 

grams 

23 

191 

2 

7 

181 

22.8 

2.5 

] 

23 

187 

2 

7 

181 

22.8 

is 

1.3 

1 

Flesh 

23 
23 

190 
192 

3 
3 

7 

7 

181 
181 

22.8 
22.8 

[    30 

2.5 

12.0 

f 

J 

23 
23 

191 
194 

1 
3 

6.93 
6.93 

181 
181 

23.1 
23.1 

|     17 

1.0 

17.0 

1 

f 

Edestin 

24 

161 

2 

6.0 

155 

19.5 

0.1 

) 

24 

158 

2 

6.0 

155 

19.5 

1  'i 

[ 

Flesh 

24 

158 

3 

6.0 

155 

19.5 

i 

24 
24 

158 
161 

1 
3 

5.94 
5.94 

156 
156 

19.8 
19.8 

t    8 

\ 

Edestin 

25 

122 

2 

5.00 

129 

16.3 

-2.0 

1 

25 

120 

2 

5.00 

129 

16.3 

-7 

-0.4 

I 

Flesh 

25 

121 

3 

5.00 

129 

16.3 

\      » 

0.8 

10.0 

[ 

25 

123 

3 

5.00 

129 

16.3 

J 

25 
25 

122 
121 

1 
3 

4.95 
4.95 

130 
130 

16.5 
16.5 

f    » 

0.8 

18.8 

t 

Edestin 

25 

122 

4 

5.00 

129 

16.3 

18 

0.5 

36.0 

Flesh 

26 

195 

3 

7.00 

179 

22.7 

-8 

1.5 

l 

Flesh 

26 

188 

3 

7.00 

179 

22.7 

-3 

1.9 

y 

26 

186 

3 

6.93 

180 

22.8 

-7 

1.2 

Edestin 

27 

138 

4 

5.50 

140 

18.1 

-10 

1.0 

! 

Flesh 

27 

138 

4 

5.50 

140 

18.1 

-7 

1.3 

27 

136 

3 

5.45 

141 

18.0 

—5 

0.5 

Edestin 

28 

124 

4 

5.50 

140 

18.1 

-14 

3.3 

\ 

Flesh 

28 

124 

3 

5.50 

140 

18.1 

—13 

3.1 

28 

123 

5 

5.45 

141 

18.0 

-11 

1.0 

Edestin 

The  flesh  ration  was  composed  as  follows:  Powdered  beef  20  parts,  sugar  25  parts,  lard 
44.5  parts,  cellulose  5  parts,  sodium  phosphate  2  parts  and  salt  mixture  3.5  parts. 

The  edestin  ration  was  composed  as  follows:  Edestin  14.93  parts,  sugar  25  parts,  lard 
46.8  parts,  cellulose  5  parts,  sodium  phosphate  3.76  parts  and  salt  mixture  3.5  parts. 

The  two  rations  had  like  calorific  value  and  like  content  of  nitrogen,  phosphorus,  fat, 
cellulose  and  salts. 

In  Table  IV,  p.  339,  we  have  a  continuance  of  the  tests  reported 
in  the  preceding  table  except  that  the  food  per  gram  of  live  weight 
of  the  edestin  rats  was  intentionally  increased  by  the  addition  of  a 
certain  amount  of  fat  to  the  ration,  the  object  being  to  determine  if 


PHOSPHORUS  METABOLISM 


339 


with  sufficient  calorific  value  the  ration  free  from  organic  phospho- 
rus would  cause  more  phosphorus  retention  than  the  ration  contain- 
ing organic  phosphorus.  Gregersen  does  not  mention  the  coinci- 
dent reduction  in  the  amount  of  food  eaten  per  gram  of  live  weight 
in  the  meat  periods.  The  food  per  gram  of  live  weight  of  the  rats 
on  the  flesh  ration  was  in  this  series  one  gram  per  28.4  grams  of  live 
weight,  while  in  the  preceding  series  this  ration  was  eaten  in  the 
amount  of  one  gram  per  25.9  grams  of  live  weight.  With  these  ad- 
vantages, together  with  those  of  influence  of  previous  feeding  and 
shorter  duration  of  the  edestin  periods,  as  previously  noted,  the  ra- 
tion which  was  free  from  organic  phosphorus  appeared  to  make  a 
better  showing  in  the  nitrogen  and  phosphorus  balances  than  did 
the  flesh  ration.  The  length  of  the  meat  period  was  in  each  case  12 
days,  during  the  last  6  of  which  balances  were  taken,  and  of  the  suc- 
ceeding edestin  period  the  average  length  was  3.7  days.  There  were, 
therefore,  four  differences  of  experimental  conditions  between  the 
two  rations  compared,  differences  as  to  calorific  value  of  the  rations, 
as  to  length  of  the  experimental  period,  as  to  advantage  from  previ- 
ous feeding,  and  as  to  nature  of  the  phosphorus  compounds  present; 
which  of  course  makes  it  impossible  to  say  what  factor  has  produced 
the  differences  in  results. 

TABLE  IV.      METABOLISM  EXPERIMENTS  ON  GROWN  RATS 

COMPARING  POWDERED  BEEF  WITH  EDESTIN 

Daily  Balances 


No. 

of  investigation, 

Length 

Food 
per  day 

Food  N 

Food  P 

N 
balance 

p 

balance 

and 

weight  of  subject 

of  period 

Ration 

in  grams 

in  days 

Milli- 

Milli- 

Milli- 

Milli- 

Grams 

grams 

grams 

grams 

grams 

29 

212 

6 

8 

268 

29.9 

-35 

-2.5 

Flesh 

29 

210 

1 

8+1 

267 

29.4 

y  33 

29 

220 

2 

8+2 

267 

29.4 

2.2 

Edestin  +  fat 

29 

220 

1 

6.8+1 

227 

24.9 

! 

29 
29 

220 
219 

2 
2 

8 
8 

268 
268 

29.9 
29.9 

+1.0 

0.1 

Flesh 

30 

307 

6 

10 

335 

37.4 

8.0 

1.5 

Flesh 

30 

313 

1 

10+2 

334 

36.7 

1 

30 

315 

2 

10+2    ' 

334 

36.7 

y  52 

4.5 

Edestin  +  fat 

30 

329 

1 

10+3 

334 

36.7 

1 

30 

336 

4 

10 

335 

37.4 

10 

1.9 

Flesh 

31 

220 

6 

8 

268 

29  .'9 

1 

-1.6 

Flesh 

31 
31 

226 
232 

1 
2 

8+1 
8+1.5 

267 
267 

29.4 
29.4 

)  » 

0.3 

Edestin  +  fat 

31 

237 

3 

8 

268 

29.9 

\- 

—1.3 

31 

238 

2 

8 

268 

29.9 

Flesh 

The  flesh  ration  was  composed  as  follows :  Powdered  beef  25  parts,  sugar  25  parts, 
lard  40  parts,  cellulose  5  parts,  sodium  phosphate  2  parts  and  salt  mixture  3  parts. 

The  edestin  ration  was  composed  as  above  except  that  edestin  and  sodium  phosphate 
replaced  the  powdered  beef,  the  nitrogen  and  phosphorus  of  the  two  rations  thus  being  held 
alike,  but  in  addition  the  edestin  ration  contained  1-3  gm.  of  lard  in  excess  of  the  amount 
present  in  the  flesh  ration. 


340 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


Table  V,  below,  sets  forth  results  of  a  comparison  of  nitrogen- 
free,  phosphorus-free  rations  with  nitrogen-free,  phosphorus-con- 
taining rations,  the  phosphorus  in  these  latter  rations  being  pres- 
ent exclusively  as  phosphates.  The  loss  of  nitrogen  and  phospho- 
rus, and  the  proportion  of  nitrogen  to  phosphorus  in  the  loss,  was 
not  affected  by  the  differences  in  these  rations. 

TABLE  V.       METABOLISM  EXPERIMENTS  WITH  MATURE  RATS  ON 
NITROGEN-FREE  RATIONS  WITH  AND  WITHOUT  PHOSPHORUS 

Daily  Balances 


No.  of  investiga- 

tion and 

weight  of 

subject 

in  grams 

32 

138 

32 

135 

32 

131 

32 

129 

32 

124 

33 

207 

33 

197 

33 

189 

33 

180 

33 

177 

34 

225 

34 

218 

34 

207 

34 

196 

34 

195 

35 

175 

35 

164 

35 

154 

35 

148 

36 

217 

36 

205 

36 

194 

36 

189 

37 

167 

37 

157 

37 

143 

37 

138 

38 

202 

38 

191 

38 


182 
171 


Length 

Food 

of 

per  day 

period 

in  days 

Grams 

3 

4 

3 

4 

3 

4 

3 

4 

4 

4 

3 

6 

3 

6 

3 

6 

3 

6 

3 

6 

3 

6 

3 

6 

3 

6 

3 

6 

3 

6 

3 

5.0 

3 

4.85 

3 

5.0 

3 

4.85 

3 

6.0 

3 

5.82 

3 

6.0 

3 

5.82 

3 

4.85 

3 

5.0 

3 

4.85 

3 

5.0 

3 

5.82 

3 

6.0 

3 

5.82 

3 

6.0 

Food  N      Food  P 


Milli- 
grams 


Milli- 
grams 


0 
0 
0 
0 
0 

0 
0 
0 
0 
0 

15.3 
15.3 
15.3 
15.3 
15.3 

12.7 
0 

12.7 
0 

15.3 
0 

15.3 
0 

0 
12.7 

0 
12.7 

0 
15.3 

0 
15.3 


N 

P 

balance 

balance 

Milli- 

Milli- 

grams 

grams 

-68 

-6.3 

-49 

-5.3 

-49 

-4.7 

-50 

-5.0 

-48 

-4.1 

-92 

-10.6 

-71 

-8.2 

-75 

—7.1 

-60 

-5.3 

-60 

-4.9 

-95 

—10.2 

-76 

-6.4 

-77 

-7.4 

-66 

—6.0 

-61 

-6.6 

-103 

-8.9 

-68 

-5.1 

-60 

-5.2 

-55 

-4.3 

-114 

—10.5 

-81 

-6.8 

-72 

-6.7 

-65 

-6.3 

-97 

-9.1 

-74 

-5.5 

-69 

—5.4 

-64 

-4.8 

-99 

-9.4 

-70 

-6.1 

-64 

-5.1 

-62 

—3.8 

N:P 

retained 
or  lost 


Ration 


10.8 
9.2 
10.4 
10.0 
11.7 

8.7 

8.7 

10.6 

11.3 

12.2 

9.3 

11.9 

10.4 

11.0 

9.2 

11.6 
13.3 
11.5 

12.8 

10.9 
11.9 
10.7 
10.3 

10.7 
13.5 
12.8 
13.3 

10.5 
11.5 
12.5 
16.3 


N-free, 
P-free 


N-free, 
P-free 


N-free, 
P-contain- 

ing 


P-containing 
P-free 

P-containing 
P-free 

P-containing 
P-free 

P-containing 
P-free 

P-free 

P-containing 
P-free 
P-containing 

P-free 

P-containing 
P-free 
P-containing 


Nitrogen-free,  phosphorus-free  ration  of  investigations  35-38  composed  as  follows:  Sugar 
50  parts,  lard  42  parts,  cellulose  5  parts  and  salt  mixture  3  parts. 

Nitrogen-free  phosphorus-containing  ration  of  investigations  35-38,  the  same  as  above 
except  for  the  addition  of   3   parts  sodium  phosphate. 

Gregersen  also  presents  results  of  a  comparison  of  nitrogen  and 
phosphorus  balances  on  phosphorus-free  rations  with  and  without 
edestin,  and,  further,  results  of  a  study  of  the  effects  of  calcium  and 
magnesium  salts  and  sodium  carbonate  added  to  the  phosphorus- 
free  edestin  ration,  on  the  elimination  of  phosphorus.  In  the  former 
experiment  the  phosphorus  loss  was  about  the  same,  whether  edes- 
tin. was  present  or  not,  there  being  no  phosphorus  retention  on  the 


PHOSPHORUS  METABOLISM  341 

nitrogen-free  ration.  In  the  latter  test  the  salts  above  mentioned 
were  also  without  influence  on  the  phosphorus  loss,  though  the 
presence  of  calcium  and  magnesium  in  the  intestine  served  to  deflect 
into  the  feces  a  considerable  portion  of  the  food  phosphorus  other- 
wise excreted  in  the  urine,  and  also  of  the  metabolic  phosphorus  of 
the  body,  as  evidenced  by  the  phosphorus  content  of  the  feces  from 
a  ration  free  from  phosphorus  but  containing  calcium  and  magne- 
sium. 

Considering  this  set  of  experiments  as  a  whole,  then,  the  trend 
of  the  evidence  suggests  organic  phosphorus  synthesis  by  rats,  but, 
as  a  demonstration  of  such  synthesis,  it  must  be  regarded  as  not  of 
conclusive  character.  As  above  noted  Gregersen  believes  that  the 
rats  synthesized  organic  from  inorganic  phosphorus  compounds,  but 
he  is  careful  not  to  claim  that  the  inorganic  compounds  are  as 
efficiently  retained  as  the  organic. 

Heubner  (1911)  conducted  feeding  experiments  on  young  dogs 
comparing  phosphates  and  lecithin  as  sources  of  phosphorus  for  the 
growing  organism.  The  dogs  had  been  kept  on  a  low-phosphorus 
diet  until  their  need  for  phosphorus  was  acute.  Lecithin  appeared 
much  superior  to  phosphates  as  a  source  of  phosphorus  for  growing 
dogs. 

Shackell  (.1911)  studied  phosphorus  metabolism  in  the  early 
cleavage  of  the  echinoderm  egg.  He  found  no  evidence  of  a  syn- 
thesis of  nuclear  material  from  alcohol-soluble  constituents  of  the 
cytoplasm  between  the  2-4  celled  stage  and  the  blastula  stage. 

Fingerling  (1912a)  demonstrated  that  ducks,  on  a  ration  which 
is  low  in  organic  phosphorus,  produce  eggs  of  normal  content  of  leci- 
thin and  nuclein  phosphorus.  He  concludes  that  the  animal  organ- 
ism possesses  the  ability  to  cover  its  requirement  of  phosphoric 
acid  for  the  formation  of  lecithin  and  nuclein  substances  just  as  eas- 
ily and  completely  with  inorganic  phosphates  as  with  organic  phos- 
phorus compounds. 

This  conclusion  could  be  justified,  in  so  positive  a  form,  only  by 
the  use  of  a  ration  free  from  organic  phosphorus,  and  by  demon- 
strating with  this  ration  that  the  original  content  of  the  body  and 
of  its  parts  was  maintained  without  loss  of  organic  phosphorus. 

J.  L.  Smith  and  W.  Mair  (1912)  studied  the  development  of 
lipoids  in  the  brain  of  the  dog.  With  reference  to  the  origin  of  the 
phosphatid,  cerebrosid  and  cholesterin  of  the  brain  they  conclude, 
from  the  very  low  content  of  the  mother's  milk  in  these  compounds, 
in  connection  with  the  considerable  daily  deposition  of  the  same  in 
the  brain  of  the  suckling  (0.045  gm.  phosphatid,  0.007  gm.  cere- 
brosid and  0.015  gm.  cholesterin),  that  these  compounds  are  syn- 
thesized from  other  substances  present  in  the  milk. 


342  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

McCollum,  Halpin  and  Drescher  (1912)  studied  lecithin  syn- 
thesis in  the  hen,  by  feeding  a  ration  which  was  very  low  in  lecithin, 
and  determining  the  lecithin  content  of  the  yolks  of  the  eggs  laid. 
The  hens  also  were  weighed.  The  hens  were  kept  from  Oct.  31  un- 
til April  15  on  a  ration  containing  30  percent  of  skim  milk  powder 
and  70  percent  of  polished  rice  which  had  been  twice  extracted  in 
boiling  alcohol  for  20-30  minutes.  From  Oct.  31  until  Jan.  30  the 
three  hens  had  gained,  respectively,  1290,  2013  and  1588  grams,  or 
33,  36.4  and  34.1  percent  in  live  weight.  They  were  assumed  not  to 
have  lost  lecithin  during  this  time,  or  later  while  laying  eggs.  Dur- 
ing the  time  from  Jan.  30  to  April  15  fifty-seven  eggs  were  laid. 
Their  yolks  contained  27.65  gm.  of  phosphorized  fats  per  hen.  The 
food  consumption  from  Oct.  31  until  March  1  averaged  58.5  grams 
per  hen  per  day.  The  amount  of  food  consumed  during  the  last 
45  of  the  70  days  during  which  the  eggs  were  laid  was  not  stated. 

The  proof  of  synthesis  of  lecithin  from  the  various  phosphor- 
ized proteins  of  the  rice  and  milk  would  be  more  complete  if  the 
authors  had  made  determinations  of  lecithin  in  the  bodies  of  the 
hens,  and  in  controls,  and  had  submitted  quantitative  determinations 
of  the  lecithin  in  the  foods,  since  they  were  not  said  to  be  entirely 
lecithin-free. 

Masslow  (1913a,  b,  c)  studied  the  biological  significance  of 
phosphorus  for  the  growing  organism  by  means  of  feeding,  metab- 
olism, tissue  analysis  and  enzyme  estimation  experiments  on  young 
dogs.  Normal  feeding  was  compared  with  feeding  on  a  phospho- 
rus-poor diet,  and  with  feeding  on  phosphorus-poor  food  plus  phos- 
phates, glycerophosphates  and  lecithin.  Casein  and  albumin  were 
also  compared. 

The  phosphorus-poor  diet  led  to  emaciation  and  finally  death, 
the  phosphorus  content  of  the  organs  having  diminished.  This  loss 
of  phosphorus  was  mainly  inorganic.  Of  the  organic  phosphorus 
only  the  lipoid  compounds  decreased.  The  brain  and  heart  appeared 
not  to  lose  phosphorus,  the  loss  being  greatest  in  liver,  intestines, 
muscles,  bone  marrow  and  kidneys.  The  ferment  functions  of  the 
organs  was  markedly  disturbed,  the  depression  of  the  action  of 
lipase,  amylase  and  diastase  being  especially  great,  there  being  also 
a  like  tendency  as  to  catalase  and  nuclease.  In  these  respects  the 
liver  suffers  most,  the  brain  and  heart  comparatively  little. 

The  addition  of  inorganic  phosphates  and  glycerophosphates 
to  the  low-phosphorus  diet  did  not  prevent  phosphorus  impoverish- 
ment. Lecithin,  however,  caused  an  enriching  of  the  organism  in 
phosphorus,  especially  in  organic  compounds  other  than  lecithin. 
The  improvement  took  place  especially  in  the  visceral  organs.  The 
ferments  were  stimulated  to  greater  activity. 


PHOSPHORUS  METABOLISM  343 

*  Exclusive  milk  diet,  maintained  beyond  the  normal  period  for 
such  food,  led  -to  disturbances  of  the  enzyme  activity  and  phospho- 
rus compounds  of  the  tissues  similar  to  those  produced  by  the  low- 
phosphorus  diet. 

Fingerling  (1913)  fed  to  goats  rations  of  straw,  blood,  nuclein, 
starch,  molasses  and  oil,  with  the  addition  of  phytin,  lecithin,  casein, 
nuclein,  nucleic  acid  or  disodium  phosphate.  No  essential  differ- 
ence was  observed  in  the  utilization  of  phosphorus  in  the  different 
forms. 

Durlach  (1913)  compared  various  phosphorus  compounds,  or- 
ganic and  inorganic,  in  feeding  and  balance  experiments  with  young 
dogs.  The  basal  ration  was  poor  in  phosphorus  and  consisted  of 
cakes  made  from  isolated  foodstuffs  and  inorganic  salts.  The  pups 
were  kept  on  the  mother's  milk  for  36-38  days ;  then  to  bring  them 
to  a  state  of  high  phosphorus  requirement  they  were  kept  on  the 
basal  ration  for  15-22  days,  after  which  time  they  were  given  the 
distinctive  diets.  Comparisons  were  made  of  monosodium  and 
monopotassium  phosphates  with  Merck's  ovo-lecithin  and  with  a 
mixture  of  monopotassium  phosphate,  lecithin,  sodium  phytate, 
casein  and  sodium  nucleate.  All  of  the  dogs  lost  in  weight.  Re- 
sults were  not  conclusive,  but  seemed  to  show  lecithin  to  possess 
superior  nutritive  value  inasmuch  as  two  of  the  three  dogs  which 
received  lecithin  were  the  only  ones  which  survived  the  experiments. 

E.  B.  Forbes  and  associates  (1914;  Ohio  Tech.  Bui.  No.  6)  have 
conducted  five  series  of  feeding,  metabolism,  and  carcass  analysis 
experiments,  involving  120  growing  swine,  in  the  comparison  of  the 
nutritive  values  of  several  organic  and  inorganic  compounds  of 
phosphorus.  The  compounds  of  interest,  in  the  usual  "chemically 
pure"  form,  were  added,  in  equivalent  amounts,  to  low-phosphorus 
standard  or  basal  rations  composed,  in  the  main,  from  comparatively 
simple  manufactured  products  of  plant  and  animal  origin.  From 
about  75  tables  of  results  four  only  are  quoted. 

Series  I,  conducted  in  April  and  May,  1908,  consisted  of  metab- 
olism experiments  with  four  pigs,  comparing  phosphates,  glycero- 
phosphates, hypophosphites,  and  nucleic  acid  from  yeast.  The 
phosphates,  glycerophosphates,  and  hypophosphites  were  mixtures, 
in  each  case,  of  salts  of  sodium,  potassium,  calcium,  magnesium  and 
iron.  The  basal  ration  consisted  of  pearl  hominy  (corn  minus  the 
skin  and  the  germ) ,  blood  albumen,  wheat  gluten,  and  salt,  with  the 
addition  of  small  amounts  of  senna  when  necessary.  The  experi- 
mental periods  were  10  days  in  length. 

That  phosphorus  in  each  form  was  absorbed  and  retained  is 
unquestionably  true,  even  the  hypophosphites  increasing  the  urin- 
ary phosphorus  and  phosphorus  retention.      Considering  normal  re- 


344 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


quirements  of  calcium  for  growth,  these  pigs  all  suffered  from  a 
deficient  intake  of  this  element.  Conditions  were  not  considered 
favorable  for  a  close  comparison  of  the  nutritive  values  of  the  phos- 
phorus compounds  involved.  The  condensed  balance  data  are  pre- 
sented in  the  following  table : 

BALANCE    EXPERIMENTS    WITH     GROWING     SWINE,     COMPARING 

PHOSPHORUS  COMPOUNDS 

Grams  per  Day — Analyses  by  H.  S.  Woods  and  A.  C.  Whittier 

Series  I;  10-day  Periods 


Pig 

and 

period 

No. 


L,ive 
weight 

Initial 
Final 

Lbs. 


Average  daily  ration 


N 

Food 
Urine 
Feces 
Bal- 
ance 


Food 
Urine 
Feces 
Bal- 
ance 


Food 
Urine 
Feces 
Bal- 
ance 


Ca 

Food 
Urine 
Feces 
Bal- 
ance 


Mg 

Food 
Urine 
Feces 
Bal- 
ance 


Food 
Urine 
Feces 
Bal- 
ance 


Period 
I 


Period 
II 


Period 
III 


Period 
I 


Period 

n 


Period 
III 


81.00 
87.75 


88.25 
99.25 


100.75 
109.00 


Hominy 1310.13 

Blood  albumen 40.25 

Wheat  gluten 40.25 

Senna 2.40 

Hominy 1314.71 

Blood  albumen 41.32 

Wheat  gluten 41.32 

Senna 1.00 

Phosphates..  (P)....    0.613 

Hominy 1245.63 

Blood  albumen 39.03 

Wheat  gluten 39.03 

Phosphates..  (P)....    0.573 

Hominy 1383.30 

Blood  albumen 41,44 

Wheat  g-luten 41.44 

Senna 2.40 

Hominy 1389.38 

Blood  albumen......    43.40 

Wheat  gluten 43.40 

Senna 2.00 

Nucleic  acid..  (P)...    0.619 

Hominy 1288.13 

Blood  albumen 40.39 

Wheat  gluten 40.39 

Nucleic  acid . .  (P) . . .    0.574 

Hominy 1451.70 

Blood  albumen 42.65 

Wheat  gluten 42.70 

Senna 2.40 

Hominy 1435.19 

Blood  albumen 44.50 

Wheat   gluten 44.50 

Senna 3.00 

Hypophosphites..(P)    0.648 

Hominy 1501.13 

Wheat  gluten 43.44 

Blood  albumen 43.44 

Senna 1.90 

Hominy 1276.66 

Blood  albumen 40.14 

Wheat  gluten 40.14 

Senna 2.00 

Glycerophosphates..    0.609 


26.421 

17.094 

1.662 

+7.665 

26.720 

17.078 

1.175 

+8.467 


25.263 

16.084 

1.150 

+8.029 


2.255 
1.169 
0.203 

+0.883 

2.272 
1.248 
0.153 

+0.871 


2.149 

1.138 
0.164 

+0.847 


0.671 

0.026 

0.309 

+0.336 

1.287 

0.435 

0.205 

+0.647 


1.210 

0.464 

0.235 

+0.511 


0.277 

0.177 

0.142 

—0.042 

0.251 

0.001 

0.074 

+0.176 


0.206 

0.005 

0.070 

+0.131 


0.287 

0.068 

0.121 

+0.098 

0.296 

0.045 

0.087 

+0.164 


0.273 

0.059 

0.102 

+0.112 


0.719 

0.164 

0.274 

+0.281 

1.114 

0.185 

0.238 

+0.691 


1.042 

0.310 

0.262 

+0.470 


Period 
I 


Period 
II 


Period 
I 


4 
Period 

n 


84.50 
92.25 


92.75 

102.50 


104.25 
112.25 


89.50 
96.50 


97.25 
109.25 


90.25 
99.75 


100.38 
107.00 


27.621 

18.405 

1.506 

+7.710 

29.292 

18.825 

1.153 

+9.314 


27.153 

17.778 

1.149 

+8.226 


2.365 

1.337 

0.198 

+0.830 

2.399 

1.370 

0.148 

+0.881 


2.223 
1.185 
0.195 

+0.843 


0.706 

0.021 

0.268 

+0.417 

1.332 

0.341 

0.217 

+0.774 


1.232 

0.562 

0.225 

+0.445 


0.286 

0.077 

0.160 

+0.049 

0.285 

0.004 

0.089 

+0.192 


0.216 

0.009 

0.073 

+0.134 


0.302 

0.078 

0.115 

+0.109 

0.343 

0.071 

0.123 

+0.149 


0.307 

0.125 

0.170 

+0.012 


28.773 
18.816 
2.083 

+7.874 

29.057 

18.237 

1.359 

+9.461 


2.471 

1.386 

0.227 

+0.858 

2.478 

1.327 

0.167 

+0.984 


0.738 

0.022 

0.262 

+0.454 

1.384 

0.630 

0.175 

+0.579 


0.303 

0.154 

0.110 

+0.039 

0.320 

0.006 

0.068 

+0.246 


0.319 

0.090 

0.125 

+0.104 

0.326 

0.061 

0.122 

+0.143 


29.563 
19.245 
2.137 

+8.181 

25.974 

18.094 

1.765 

+6.115 


2.545 

1.360 

0.229 

+0.956 

2.208 

1.349 

0.220 

+0.639 


0.760 

0.019 

0.344 

+0.397 

1.265 

0.225 

0.368 

+0.672 


0.287 

0.162 

0.184 

-0.059 

0.295 

0.008 

0.140 

+0.147 


0.323 

0.096 

0.125 

+0.102 

0.575 

0.119 

0.236 

-r  0.220 


0.754 

0.132 

0.226 

+0.396 

0.814 

0.298 

0.234 

+0.282 


0.739 

0.389 

0.212 

+0.138 


0.788 

0.178 

0.194 

+0.416 

1.063 

0.173 

0.197 

+0.693 


0.809 

0.131 

0.493 

+0.185 

1.138 

0.116 

0.330 

+0.692 


PHOSPHORUS  METABOLISM  345 

Series  II,  conducted  in  November  and  December  1908,  consist- 
ed of  a  feeding  and  carcass  analysis  experiment,  covering  56  days, 
and  involving  the  use  of  30  pigs,  five  individuals  in  each  of  six  lots. 
The  basal  ration  was  composed  of  pearl  hominy,  blood  albumen, 
wheat  gluten,  and  corn  bran.  Salt  was  allowed  ad  libitum.  Corn 
meal  and  chalk  were  introduced  into  the  diet  at  certain  times,  for 
corrective  purposes.  The  phosphorus  compounds  compared  were 
the  same  as  in  Series  I.  The  several  lots  received  rations  as  fol- 
lows: 

Lot  1,  basal  ration  plus  hypophosphites 

Lot  2,  basal  ration  plus  hypophosphites  and  nucleic  acid 

Lot  3,  basal  ration  plus  glycerophosphates 

Lot  4,  basal  ration  plus  phosphates 

Lot  5,  basal  ration  alone 

Lot  6,  check  lot,  killed  at  beginning  of  experiment 

The  pigs  in  lots  1  and  2  suffered  from  great  weakness  and 
lameness;  they  were  also  subject  to  indigestion.  The  legs  were  weak, 
especially  the  hind  ones,  which  trembled,  and  bowed  out,  and  the 
feet  were  set  far  under  the  body.  The  fore  feet  were  also  sore.  The 
muscular  control  was  poor,  and  difficulty  was  experienced  in  step- 
ping up  6  inches  onto  the  feeding  platform.  These  pigs  lay  down 
most  of  the  time,  and  if  disturbed  would  at  once  lie  down  again. 
They  ate  well,  but  moved  slowly,  carefully  and  without  spirit. 
Lot  5,  which  received  no  phosphorus  supplement,  behaved  much  as 
did  lots  1  and  2,  but  the  abnormal  tendencies  were  less  pronounced. 
Lot  3,  receiving  glycerophosphates,  were  entirely  normal ;  they  ran 
and. played  in  the  best  of  spirits.  Lot  4,  receiving  phosphates,  also 
remained  in  good  condition,  though  they  were  less  active  than  the 
pigs  receiving  glycerophosphates. 

Considerable  difficulty  was  experienced  in  the  management  of 
the  feeding,  the  lots  receiving  hypophosphites  and  nucleic  acid  re- 
quiring frequent  reduction  in  the  amount  of  food  given,  and  in  the 
amount  of  the  phosphorus  supplements.  These  reductions  it  was 
not  possible  to  make  up  by  subsequent  increase.  On  these  accounts 
the  food  consumption  could  not  be  maintained  uniform  in  all  lots,  a 
fact  which  resulted  in  certain  advantage  to  the  pigs  receiving 
glycerophosphates.  The  nutritional  disorders  in  certain  lots,  as 
above  noted,  were  successfully  combatted  by  the  feeding  of  limit- 
ed amounts  of  corn,  which  in  this  relation  appeared  to  possess 
marked  curative  value,  perhaps  due  to  its  organic  phosphorus  com- 
pounds— perhaps  to  vitamines.  The  basal  ration,  by  the  way,  was 
very  poor  in  fats. 


346  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Conclusions  as  to  the  effects  of  the  phosphatic  supplements 
were  drawn  from  slaughter  weights  of  parts  and  organs,  and  an- 
alyses of  the  hams,  brains,  livers,  kidneys,  femora,  and  tibiae,  the 
data  in  all  cases  representing  composite  samples  from  the  5  indi- 
viduals in  an  experimental  lot.  For  a  discussion  of  the  results  see 
p.  352. 

Series  III,  conducted  during  November  and  December  1909  and 
January  1910,  was  in  every  way  similar  to  Series  II.  The  feeding 
covered  a  period  of  70  days,  and  the  experiment  involved  35  pigs  in 
7  lots  of  5  each.  The  basal  ration  consisted  of  pearl  hominy,  wheat 
gluten,  blood  albumen,  and  corn  bran.  Salt  was  allowed  ad  libitum. 
To  this  basal  ration  were  added,  at  various  times  during  the  ex- 
periment, chalk,  soda,  sugar  and  corn  meal,  in  efforts  to  relieve  dif- 
ficulties in  the  feeding.  Without  some  corn  it  seemed  impossible 
to  keep  the  pigs  in  condition  for  experimentation.  The  phospho- 
rus compounds  fed  were  the  same  mixtures  used  in  Series  I  and 
II.      The  several  lots  received  rations  as  follows : 

Lot  1,  basal  ration  plus  hypophosphites 

Lot  2,  basal  ration  plus  nucleic  acid 

Lot  3,  basal  ration.plus  glycerophosphates 

Lot  4,  basal  ration  plus  phosphates 

Lot  5,  basal  ration  plus  phytin 

Lot  6,  basal  ration  alone 

Lot  7,  check  lot,  killed  at  beginning  of  experiment. 

Difficulties  in  the  feeding  were  encountered  in  all  lots  except 
the  one  receiving  glycerophosphates,  and  the  one  which  received  no 
phosphorus  supplement.  The  pigs  receiving  glycerophosphates 
were  much  the  most  spirited  and  active  of  any  in  the  series.  Hypo- 
phosphites  and  nucleic  acid,  when  fed  in  amounts  furnishing  2 
grams  of  phosphorus  daily  to  each  lot  of  5  pigs  caused  nausea,  vom- 
iting, and  indigestion.  Phytin  appeared  to  cause  indigestion,  but 
not  pronounced  nausea.  As  a  whole  the  pigs  in  this  series  were 
much  less  tolerant  of  the  phosphorus  compounds  fed  (other  than 
glycerophosphates)  than  those  in  Series  II.  In  this  experiment 
even  the  pigs  receiving  orthophosphates  exhibited  very  limited  tol- 
erance for  the  mineral  supplement. 

Series  IV,  conducted  in  November  and  December  1910  and  Jan- 
uary 1911,  was  similar  in  method  to  Series  II  and  III,  the  feeding 
extending  over  a  period  of  70  days,  and  the  experiment  involving  45 
pigs.  The  basal  ration  in  lots  1-5  consisted  of  pearl  hominy,  wheat 
gluten,  blood  albumen,  corn  bran  and  agar-agar,  salt  being  allowed 


PHOSPHORUS  METABOLISM  347 

ad  libitum.  To  this  ration  were  added,  during  portions  of  the  ex- 
periment, mangel  wurzels  and  potassium  citrate,  for  corrective  pur- 
poses. Lots  6-8  received  a  similar  ration,  except  that  corn  replaced 
the  pearl  hominy  used  in  the  food  for  lots  1-5.  The  hypophosphites, 
nucleic  acid  and  phosphates  used  were  mostly  of  the  same  lots  as 
those  used  in  the  earlier  series,  but  the  glycerophosphate  mixture 
was  soon  exhausted  and  was  then  replaced  by  calcium  glycerophos- 
phate alone.  The  phytin  was  in  part  a  commercial  product,  and  in 
part  a  preparation  of  the  related  compound  from  wheat  bran.  The 
several  lots  received  rations  as  follows : 

Lot  1,  hominy  basal  ration  alone 

Lot  2,  hominy  basal  ration  plus  nucleic  acid 

Lot  3,  hominy  basal  ration  plus  glycerophosphates 

Lot  4,  hominy  basal  ration  plus  phosphates 

Lot  5,  hominy  basal  ration  plus  phytin 

Lot  6,  corn  basal  ration  plus  precipitated  bone  flour 

Lot  7,  corn  basal  ration  plus  glycerophosphates 

Lot  8,  corn  basal  ration  alone 

Lot  9,  check  lot ;  killed  at  beginning  of  experiment. 

The  pigs  in  this  experiment  exhibited,  in  general,  the  same 
symptoms  as  noted  with  regard  to  the  previous  series.  The  pigs 
which  received  nucleic  acid  and  phytin,  and  those  on  the  hominy 
basal  ration  alone,  suffered  from  much  weakness  and  soreness  of  the 
feet  and  legs.  There  was  also  considerable  trouble  with  indigestion 
in  these  lots,  as  also  in  the  phosphate  lot,  and  the  pigs  receiving 
phytin  showed  some  appearance  of  nausea.  The  glycerophosphate 
pigs,  as  usual,  were  entirely  normal,  and  suffered  only  from  the 
restricted  food  allowance  necessitated  by  the  unsatisfactory  behav- 
ior of  other  lots.  Lots  1,  2,  4  and  5  ate  their  own  feces ;  lots  3,  6,  7 
and  8  showed  no  such  tendency. 

In  Series  II,  III  and  IV  it  was  necessary,  because  of  digestive 
disturbance  or  lack  of  appetite,  to  reduce  the  amount  of  food  allowed 
to  one  or  more  of  the  experimental  lots  on  51  dates.  The  pigs 
receiving  glycerophosphates  were  not  the  occasion  for  any  one  of 
these  food  reductions,  their  immunity  to  nutritional  disturbance  be- 
ing complete,  and  in  marked  contrast  to  the  behavior  of  all  other 
lots.  Each  of  the  other  rations  contributed  in  somewhat  nearly  the 
same  proportion  to  the  occasions  for  reduction  of  food  allowance. 

Series  V,  conducted  during  March,  April,  May  and  June  1913, 
was  a  carefully  controlled  set  of  balance  experiments  comparing 
glycerophosphates  and  phosphates,  and  was  terminated  by  a  slaugh- 
ter test  and  a  complete  chemical  accounting  for  the  bodies  of  the 
experimental  subjects.      Six  barrows,  all  of  the  same  litter,  were 


348    •  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

used  in  this  investigation.  Two  were  killed  as  controls  at  the  be- 
ginning of  the  study,  the  other  four  serving  as  subjects  for  the 
metabolism  experiments.  With  the  same  low-phosphorus  basal  ra- 
tion, two  pigs  received  phosphates,  and  two  glycerophosphates,  the 
mineral  bases  in  these  supplements  being  proportioned,  one  to  an- 
other, as  in  sow's  milk.  The  basal  ration  was  composed,  as  in  the 
earlier  studies,  from  pearl  hominy,  blood  albumen,  wheat  gluten, 
corn  bran,  agar-agar  and  salt. 

As  in  the  previous  work,  much  difficulty  was  experienced  in 
the  feeding  of  the  pigs,  other  than  those  receiving  glycerophos- 
phates, and  frequent  readjustment  of  the  level  of  intake  of  food  was 
necessary.  Very  fine  grinding,  and  extensive  dilution  of  the  food 
with  water,  were  tried  in  efforts  to  solve  the  problem  of  feeding  the 
orthophosphates,  but  without  distinct  success.  The  experiment 
finally  terminated  itself  in  the  digestive  collapse  of  one  of  the  pigs 
receiving  phosphates. 

A  part  of  the  balance  data,  reduced  to  amounts  per  kilogram  of 
live  weight,  are  presented  in  the  following  tables : 


PHOSPHOEUS  METABOLISM 


S49 


I" 


^    ^       v    hi 


v 


oooo     oooo 

OOPM        OOh-tO 


OOOO      oooo 

HOOtO        OOHtO 
CJlOiOSUl        G0--JOCJ1 


OOOO   OOOO 

oooo  oooo 

COOOOOi       O0tOt—O5 


hj     H        .^     H 


11  H, 


OOOO      oooo 

OCMtO        OOH-CO 


OOOO        OOOO 
h-OOW        OOHtO 


W  —  2.S 
"*3 


P  <! 

3  n> 


3±.P 
ft  P; 

S  ^ 


p  £  So  ^ 

p  8  5  g,  p 
to 


p  ^dw 
p"8-.o 


p  ^dbj 


OOOO   oooo 
oooo  oooo 

COtOH-*Oi   CCCOh-O^l 


oooo  oooo 
oooo  oooo 


oooo     oooo 

I— »OHOJ        HOMW 


OOOO  OOOO 
h-OOCO  I— '  O  CO  tO 
OOtOOitNS        M-tCUUO 


OOOO      oooo 
oocooo     ootow 

o\coii      ffioxoi 


OOOO      oooo 
oooo>      OOOO 

OJI-'tO-J        OOF—tO-J 


OOOO      oooo 

h^OI— *CC        h-O^-OO 
02H*-05~a       050105-q 


OOOO  OOOO 
HOOtO  HOOM 
OCCOCDI— '        h£-02COCO 


P*lCJ*J 

Bag*" 

CD 


P  ^d^l  i=< 

J-*  O)   i-j   o  & 

P  O  2.  o  R 

B»3pi 


w 

P    *id*l 

P   <T    P    P* 


p  3d*i 

pRa.g  ^ 

3»PS, 
2  m  ffl 


w 

M 

►  2 

*  S 

o  g 

^  § 

r>  H 

l> 

Kj  S| 

5 

O   H 

H     ^2 

o  ^ 

CO 

►fl      HH 

O 

n  W 

m 

^ 

•=j 

02     -. 

n 

hj    O 

a' 

03 

<5 

td 

f 

>  2 

S. 
S" 

p 

3 

o 
M 

1     * 

i-3 

HH 

H> 

l-H 

^ 

1 

21    M 

e 

cT 

on- 

d 

p 

3 

w  2 

p 

Ml 

►  w 

CO 

c-1 

M 

© 

H   S| 

•^ 

w 

W  "^ 

p 

3 

5  ° 
5  3 

^<oo<o  <0>CO^<0 
oototo  OOOJW 
tooco-g     if-oi- 'Cji 


toooco     toooco 

h- *)— »00t— *        PM<IO 

o*Jvjo     oococnoo 


tOOOOJ        BOOM 


cotoooo 


CO  to  Ol  00 


p  "d^ 

gag* 


w 

p  ^)d>rj 

pR2.8  « 


CO 


S50 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


hj    hj       hj    hj 


~,         *d      m, 


oooo     oooo 

OOHKJ        OOtOH 


OOOO        OOOO 

MOOtO        h-OOtO 


OOOO   OOOO 
OOOO   OOOO 

t^toocn      west— m 


oooo  OOOO 
OOOO  OOOO 
K^i— it>o»q       COt— *tC05 


oooo      OOOO 
>— 'Ol— ^rf*       I— 'Oil— i  to 


hd    h9       hj    h 


OOOO  oooo 
OOHM  OOOtO 
tC4-~-»JCO       CCr.GCOJ 


OOOO        OOOO 
HOCtO        I— 'OOtO 


OOOO   OOOO 
OOOO   OOOO 


OOOO   OOOO 
OOOO   OOOO 


fci^ 


p. 

>■ 

p 

< 

3 

n 

p. 

p 

«J 

«3 

0) 

Tl 

p. 

p 

15" 


P  *jej*] 
o  al  a 


W 

P  •TjCl'T)  .. 
3  (D  3  p* 


P  ^dw 
SSSs  p 


P  ^<=l*Tj  _. 

3  8  3  £*) 


oooo  oooo 

h- 'O^-'CO   h--0*-*^ 


p  ^c|w 

pS2.o 
3  n>  3  2 
o  to  (t  P" 


OOOO  OOOO 
k-OOtO  h-OOlO 
CntOOSlf*      tococxo 


OOOO        OOOO 

ooww      o  o  to  OJ 

CDOtOOO        W^OOOtO 


IOOOCO        POOM 
OQ0O00      Gil—  00*. 


OOOO        OOOO 
HOOtO        I— 'OOtO 

tocooso     oicnto*. 


w 

3§B-8 


OOOO        OOOO 

ootceo      ootow 

QlO-^JtO        CDOCDQ0 


pooto     rooooo 

CDtO-JGO        CCIO--ICO 


P  ^CJ1^ 
CD 


td 

P  ^cJ1^ 

£8  2.8  a 


2.  S 
ft    •  ' 

*    td 

ft    i6 

5*  P 
p- 

3^ 

o   j£ 

a  N 

p    p 
«4 


© 

op. 

p 
0 


2  § 

O  H 

o  ^ 

t*  £  o 

8  a  o 

H  2 


w 

I— I 

r 

O 

o 
> 


PHOSPHORUS  METABOLISM 


851 


Jti    ^        V    H 


h3    ^ 


^     1-9 


oooo     oooo 

OOMtO        OOOi—» 
■  -J!— KOQt- »       «*4)— '0005 


oooo      oooo 

OKQtO        OOOI— » 
COH- 'QIC?!       O^GCCRCC 


8^ 


si  ^ 

(K)  3  trq  • 

IT" 


p  < 
3  a 

p."1 


B<« 


OOOO        OOOO 

p  o  o  ^      o  o  o  ro 


oooo 

OOO— ' 


oooo 

HOOM 
COOOh^Oi 


oooo   OOOO 

oooo  oooo 

COtOOOi        tOtOOrf^ 


oooo  oooo 
oooo  oooo 
ostoiss-j     ton-,  tooi 


oooo  oooo 
homo:  ^-  o  *-  ro 
Cioioooo     towwoo 


oooo     oooo 

l—OOCO        OOO'-' 
COtf».~JtO        O0OJO5O5 


OOOO      oooo 

OOOO        OOOO 


p  $3g 


SjJSft 


8.  p 


p  *lQw 

J7*  re    I")    O      HH 


P  ^c^w 
2  m  » 


oooo  oooo 
oooo  oooo 


oooo     oooo 
oojcooi     ^jSKoo 


OOOO        OOOO 
POO>— i        >— 'OOtO 


OOOO       OOOO 

ootoco      oowto 

03000  01       OOM-J 


MOOU  h-"OObO 
t— 'hO^JI— »  Oil— 'O^CO 
C0OC0  00       (OfflOUl 


oooo     oooo 
ooccto      oo  5 

W-OOH^        OOOCOOT 


t2SE5       MOOU 

oconoto     »jaao 


P   ^<3w    « 

p  S  -o  S 


p  ^c^w 
pRS.g 

,reg£ 


3  <TJ  B  2 


p  ^cjw 


p  o 


- 


P  ^q« 
o  o  a  P* 


p  IQh 

sBS-8  a 


w>    W   >-( 


©   2. 


H 


352  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Discussion  of  Results.  From  the  results  of  Series  I  it  is  mani- 
fest that  orthophosphates,  hypophosphites  and  yeast  nucleic  acid 
may  all  be  absorbed  by  swine,  and  may  be  retained  in  considerable 
quantity  for  at  least  10  days.  That  the  retention  of  each  of  these 
compounds  may  be  permanent  seems  altogether  probable.  In  the 
case  of  hypophosphites  this  would  involve  a  further  oxidation  of 
the  phosphorus  to  the  ortho  form. 

Under  the  conditions  of  Series  II,  III  and  IV,  there  was  some 
evidence  to  suggest  nutritive  superiority  of  glycerophosphates 
to  orthophosphates,  nucleic  acid,  phytin  and  hypophosphites,  es- 
pecially in  relation  to  the  proportion  of  muscular  tissue  and  fat  in 
the  increase,  and  the  breaking  strength  and  ash  per  cubic  centimeter 
of  volume  of  the  bones,  but  the  evidence  was  not  sufficiently  of  one 
sort  to  establish  facts  with  certainty. 

It  seems  quite  possible  that  the  amount  of  exercise  taken  by 
the  pigs,  as  determined  simply  by  the  state  of  feeling  induced  by 
the  phosphorus  compounds  fed,  entirely  irrespective  of  fundamental 
nutritive  effects,  may  have  entered  prominently  into  the  determina- 
tion of  the  relative  development  and  even  the  composition  of  the 
parts. 

The  final  series  of  experiments,  No.  5,  was  intended  to  establish 
any  such  fundamental  differences  as  there  may  be  in  the  nutritive 
values  of  phosphates  and  glycerophosphates.  It  included  complete 
and  protracted  mineral  and  nitrogen  balances,  complete  chemical  ac- 
counting for  the  carcasses  of  the  pigs,  determinations  of  digestibil- 
ity of  the  food  constituents,  determinations  of  the  comparative  de- 
velopment and  composition  of  various  organs  and  tissues,  and  daily 
determinations  of  the  nitrogen,  creatinin  and  ammonia  of  the  urine. 

Throughout  this  exhaustive  study  there  was  marked  similar- 
ity in  the  results  from  both  rations.  Aside  from  the  much  greater 
acceptability  of  the  glycerophosphates  than  the  phosphates  there 
were  no  differences  of  note  in  results  from  the  two  pairs  of  pigs. 

The  result  of  this  series  was  a  remarkably  uniform  and  consist- 
ent set  of  observations  of  many  sorts  indicating  that,  at  least  under 
the  artificial  conditions  of  this  experiment,  there  are  no  essential 
differences  in  the  fundamental  nutritive  value  of  phosphates  and 
glycerophosphates  for  purposes  of  growth  in  swine. 

Conclusions.  Phosphates,  hypophosphites,  glycerophosphates, 
nucleic  acid  (from  yeast)  and  phytin  are  all  absorbed  and  retained, 
and  apparently  utilized,  by  growing  swine,  when  added  in  the  pure 
form  to  rations  low  in  phosphorus  but  capable  of  maintaining  phos- 
phorus equilibrium. 


PHOSPHORUS  METABOLISM  353 

Marked  differences  were  observed  in  the  tolerance  of  swine 
toward  the  several  phosphorus  compounds  used.  These  were  found 
to  compare  as  to  acceptability  to  swine,  when  fed  in  quantities  fur- 
nishing equal  amounts  of  phosphorus,  in  the  following  order,  the 
most  acceptable  being  mentioned  first,  and  the  others  in  order  of  de- 
creasing acceptability :  glycerophosphates,  orthophosphates,  phytin, 
nucleic  acid  and  hypophosphites. 

That  the  differences  in  acceptability  of  these  isolated  compounds 
similarly  affect  the  foodstuffs  in  which  they  are  naturally  combined 
seems  certainly  not  to  be  a  fact.  These  differences  in  acceptability 
were  not  found  definitely  related  to  more  fundamental  nutritive 
effects. 

That  the  particular  organic  compounds  used  in  this  investiga- 
tion (nucleic  acid,  phytin  and  glycerophosphates)  have  nutritive 
values,  to  growing  swine,  superior  to  the  inorganic  compounds  used 
(orthophosphates  and  hypophosphites)  was  not  shown.  No  funda- 
mental differences  in  the  nutritive  values  of  the  phosphorus  com- 
pounds studied  were  established. 

No  basis,  therefore,  was  discovered  for  a  differentiation  be- 
tween the  nutritive  values  of  organic  and  inorganic  phosphorus  com- 
pounds generally.  It  should  be  borne  in  mind,  however,  that  no 
representatives  of  the  two  classes,  phosphoproteins  and  lecithins, 
were  included  in  this  investigation,  and  results  obtained  under  con- 
ditions of  such  rigid  experimental  control  may  not  accurately  rep- 
resent the  facts  under  optimum  normal  conditions  of  life.  These 
results  are  not  considered  to  controvert  evidence  as  to  specific  ther- 
apeutic effects  of  the  same  compounds  in  relations  other  than  those 
considered  in  this  study. 

The  amount  of  phosphorus  which  an  animal  will  tolerate,  when 
added  to  the  ration  in  readily  soluble  form,  is  definitely  limited  at  an 
amount  much  less  than  that  which  will  be  acceptable  in  its  natural 
relationships  in  foods. 

From  the  great  difficulty  experienced  in  the  feeding  of  yeast 
nucleic  acid  and  of  commercial  phytin,  as  well  as  the  related  com- 
pound from  wheat  bran,  it  is  concluded  that  the  isolation  of  such 
compounds  from  natural  products  alters  at  least  their  therapeutic 
effects  in  such  manner  that  it  becomes  impossible  to  state,  from  in- 
vestigations of  this  sort,  on  pure  compounds,  what  may  be  their 
nutritive  values  in  their  natural  relationships  in  common  foods. 

It  seems  unlikely  that,  with  growing  animals,  any  ration  com- 
posed from  natural  foods,  and  supplying  the  nitrogen  requirement, 
will  fail  to  furnish  enough  phosphorus  to  maintain  phosphorus  equi- 


S54  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

librium.  That  many  rations  compounded  from  common  foods  are 
lacking  in  the  amount  of  phosphorus  essential  to  maximum  reten- 
tion and  growth,  however,  is  as  certainly  true. 

The  addition  of  comparatively  small  amounts  of  corn  to  rations 
compounded  from  simple  manufactured  products  of  plant  and  ani- 
mal origin  may  enhance  the  nutritive  value  of  such  rations  to  an  ex- 
tent out  of  proportion  to  the  amount  of  corn  added,  and  may  affect 
the  results  of  such  comparisons  of  phosphorus  compounds  as  are 
here  reported  in  definite  and  fundamental  ways  determined  by  the 
state  of  nutrition  of  the  animal,  especially  with  reference  to  nutri- 
tive reserves,  resulting  from  the  previous  feeding  and  life  activity. 

In  spite  of  a  marked  tendency  toward  constancy  of  composition, 
appreciable  differences  were  noted  in  the  chemical  constitution  of 
the  organs  and  tissues  of  different  experimental  lots  of  swine,  these 
differences  apparently  being  due  in  part  to  the  phosphorus  com- 
pounds fed,  and  in  part  to  other  factors  which  were  not  controlled. 

Such  differences  in  the  composition  of  tissues  as  were  observed 
might  be  accounted  for  as  due  to  variations  in  the  liquid  content  of 
the  parts,  the  salts  varying  accordingly,  or  to  the  composition  of 
supporting  structures  or  unorganized  nutritive  materials. 


There  is  also,  in  medical  iiterature,  a  vast  amount  of  clinical  evi- 
dence to  which  we  could  refer,  much  of  it  flimsy,  it  is  true,  but 
other  portions  of  undoubted  value  as  showing  specific  effects  of 
lecithin  which  are  not  possessed  by  phosphates,  especially  the 
direct  contribution  of  lecithin  to  the  lecithin  content  of  the  blood 
serum  and  tissues,  and  to  the  increase  of  the  red  blood  corpuscles. 

See  summary,  p.  534. 

.    ORGANIC  PHOSPHORUS  SYNTHESIS  IN  FASTING  SALMON 

This  problem  of  synthesis  of  organic  phosphorus  compounds 
has  also  been  attacked  from  another  angle  in  the  study  of  the  com- 
position of  salmon  at  different  stages  of  sexual  maturity,  these 
fishes,  in  the  journey  to  their  spawning  grounds,  traveling  long  dis- 
tances, apparently  without  taking  food,  and  at  the  same  time 
accomplishing  the  organization  of  great  masses  of  the  sexual  ele- 
ments. 

Miescher  (1881),  in  studying  the  life  of  the  Rhine  salmon,  came 
to  the  conclusion  that  the  very  great  development  of  nuclear 
material  in  the  sexual  elements  during  fast  took  place  at  the  expense 
of  the  muscular  system.    This  investigation  was  not  concluded. 

Paton  et  al.  (1897-8,  1898)  made  an  extensive  series  of  observa- 
tions on  the  metabolism  of  the  salmon  during  its  life  in  fresh  water. 


PHOSPHORUS  METABOLISM  355 

Lecithin  was  determined  from  the  ether  extract;  inorganic  phos- 
phorus by  extraction  of  the  ether-extraction  residue  with  0.2  per- 
cent hydrochloric  acid,  while  the  phosphorus  in  nuclein  and  pseudo- 
nuclein  was  determined  in  the  residue  from  the  above-mentioned 
hydrochloric  acid  extraction. 

The  matter  of  especial  interest  in  this  investigation  is  the  evi- 
dence on  the  subject  of  synthesis  of  organic  phosphorus  compounds. 
By  tissue  analysis  of  fishes  from  the  estuary,  and  later  from  the 
upper  river  waters,  after  the  development  of  the  ovaries  and  testes 
during  fast,  conclusions  were  drawn  as  to  the  transformation  of 
phosphorus  compounds  within  the  body.  In  the  female  fish  the 
lecithin  of  the  trunk  muscles  was  found  to  be  insufficient  to  yield 
the  lecithin  phosphorus  of  the  ovaries,  the  total  phosphorus  lost  by 
the  trunk  muscles  being  only  just  sufficient  to  yield  the  total  phos- 
phorus laid  on  by  the  ovaries;  but  in  the  male  fish  more  lecithin 
is  lost  from  the  muscles  than  is -required  by  the  testes. 

The  authors  concluded  that  the  phosphorus  stored  in  the 
muscles  as  simple  phosphates  is  transferred  to  the  ovaries  and 
testes,  and  there  built  up  into  organic  combinations,  lecithin  consti- 
tuting an  important  step  in  the  conversion  of  the  phosphates  into 
true  nuclein  in  the  testes,  and  into  the  intermediate  pseudonuclein, 
ichthulin,  and,  later,  true  nuclein  in  the  ovaries. 

The  study  of  this  problem  is  attended  by  difficulties  which 
warrant  the  suspension  of  our  judgment  as  to  the  significance  of 
the  results  at  least  until  there  shall  have  been  made  complete  chem- 
ical accountings  for  the  entire  bodies  of  the  fishes. 

Milroy  (1908)  has  made  similar  studies  on  the  herring,  but 
definite  conclusions  were  not  reached  as  to  the  source  of  the  phos- 
phorus compounds  of  the  ovaries  and  testes. 

ADVANTAGE  OF  COMPLEXITY  OF  ORGANIZATION  IN  FOOD 

PHOSPHORUS 

On  a  priori  grounds  a  certain  superiority  of  the  organic  as  com- 
pared with  the  inorganic  phosphorus  compounds  would  be  consid- 
ered to  be  due  simply  to  the  greater  complexity  of  organization, 
since  in  the  case  of  those  compounds  (the  lecithins  and  the  nucleins) 
which  are  absorbed  in  part  as  such,  or  in  a  state  of  incomplete 
digestive  cleavage,  a  certain  amount  of  synthetic  activity  is  prob- 
ably spared  the  animal.  As  to  how  important  a  matter  this  may  be 
we  can  only  speculate.  With  reference  to  nitrogen  compounds,  how- 
ever, the  maintenance  of  nitrogen  equilibrium  with  the  less  com- 
pletely split  proteins  seems  to  be  appreciably  more  efficient  than 


556  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

with  the  products  of  complete  digestive  cleavage,  and  the  same 
principle  may  be  operative  in  the  case  of  some  of  the  organic  phos- 
phorus compounds. 

Abderhalden  and  Rona  (1904)  found,  in  experiments  with  mice, 
that  pancreatic  digestion  products  of  casein  are  about  as  valuable 
for  maintenance  of  live  weight  as  casein  itself,  that  the  further  split 
products  of  peptic-pancreatic  digestion  of  casein  are  of  less  value, 
though  of  some  use,  while  the  products!  of  sulphuric  acid  hydrolysis 
seemed  to  be  without  value.  These  authors  (1905)  concluded  that 
a  dog  was  able  to  utilize  pancreatic  digestion  products  of  casein, 
these  products  consisting  of  amino  acids  and  complex  sub- 
stances not  giving  the  biuret  reaction;  but  the  products  of 
acid  hydrolysis  of  casein  were  not  able  to  maintain  nitro- 
gen equilibrium.  In  later  work,  however,  Abderhalden  (1912) 
showed,  in  protracted  nitrogen  balance  experiments,  that  dogs 
are  able  to  maintain  nitrogen  equilibrium  and  retention 
on  products  of  either  acid  or  enzymatic  splitting  of  proteins, 
or  on  a  mixture  of  recognized  amino-acids.  Abderhalden  expresses 
the  belief  that  the  organism  must  have  prepared  for  itself  all 
of  the  building  stones  which  are  specific  for  phosphatids,  since 
these  were  not  fed.  To  have  demonstrated  this  point,  however, 
would  have  required  carcass  analyses,  which  were  not  a  part  of  this 
investigation. 

D.  D.  Van  Slyke  and  G.  F.  White  (1911),  in  protein  digestion  and 
retention  experiments  with  dogs,  found  that  with  various  meats  the 
rate  of  digestion  and  absorption,  as  indicated  by  the  rate  of  elim- 
ination, was,  in  general,  the  reverse  of  the  amount  of  nitrogen 
retained  from  each  at  the  end  of  24  hours. 

In  explanation  of  these  results  the  authors  (Van  Slyke  and 
White)  suggest  that  a  larger  proportion  of  the  more  readily  digested 
proteins  may  be  absorbed  in  the  form  of  the  lowest  cleavage  prod- 
ucts, which  Carrel,  Levene,  Meyer  and  Manson,  and  also  Voit  and 
Zisterer  and  others,  have  shown  to  be  less  capable  than  the  higher 
cleavage  products  to  maintain  nitrogenous  equilibrium. 

In  the  light  of  these  experiments  it  is  quite  conceivable  that, 
other  things  being  equal,  some  of  the  more  highly  organized  organic 
phosphorus  compounds  should  possess,  on  account  of  their  state 
of  organization,  nutritive  value  greater  than  that  of  the  simplest 
inorganic  phosphates. 

A  further  advantage  to  the  animal  in  having  the  phosphorus 
compounds  of  the  food  organically  combined  is  suggested  by  the 
experience  of  one  of  us  (E.  B.  F.)  in  the  feeding  of  pure  phosphates 
along  with  a  low-phosphorus  basal  ration  to  swine.  The  ready  solu- 


PHOSPHORUS  METABOLISM  357 

bility  of  the  uncombined  phosphates  results  in  such  a  concentration 
of  these  salts  in  the  digestive  tract  as  causes  nausea  or  catharsis. 
Much  larger  amounts  of  phosphorus  may  be  utilized  in  a  normal 
manner  if  they  are  gradually  liberated  in  the  usual  way  by  the 
digestive  cleavage  of  the  organic  complexes  with  which  they  are 
combined. 

FEEDING  EXPERIMENTS  WITH  RATS  AND  MICE  ON  RATIONS  COM- 
POSED OF  SIMPLE  PURIFIED  NUTRIENTS 

An  important  addition  to  the  evidence  on  organic  phosphorus 
synthesis  has  been  made  through  a  number  of  extensive  series  of 
feeding  experiments  with  rats  and  mice,  most  of  them  having  for 
their  object  especially  the  study  of  synthesis  of  nitrogen  compounds 
in  general,  by  experiments  on  growth  and  reproduction,  on  rations 
of  simple  purified  nutrients. 

Rohmann  conducted  experiments  with  mice  which  show  that 
even  though  phosphoproteins  and  nucleoproteins  be  present  in  abun- 
dance, and  mineral  salts  be  furnished  as  phosphates,  nitrates,  chlo- 
rides and  lactates  of  the  more  important  bases  in  the  body,  there 
may  still  be  nutritive  deficiencies  in  the  ration  such  as  gradually, 
through  the  course  of  successive  generations,  make  growth  impos- 
sible. 

Rohmann  (1902,  1907)  successfully  reared  two  generations  of 
mice  on  an  artificial  food  mixture  containing  its  phosphorus  as  phos- 
phoprotein,  nucleoprotein  and  phosphates,  but  failed  to  rear  the 
third  generation.  In  a  later  paper  (1908)  Rohmann  reported  suc- 
cess in  rearing  three  generations  of  mice  to  maturity  on  artificial 
foods,  but  the  fourth  generation  could  not  be  reared.  The  nutrients 
used  in  these  experiments  were  casein,  egg  albumin,  nucleoprotein 
from  liver,  potato  starch,  wheat  starch,  margarine,  malt,  chicken 
protein  and  a  salt  mixture  compounded  as  follows : 

10  parts  calcium  phosphate 

40  parts  acid  potassium  phosphate 

20  parts  sodium  chloride 

15  parts  sodium  citrate 
8  parts  magnesium  citrate 
8  parts  calcium  lactate 

Mendel  suggested  that  Rohmann's  final  failure  was  due  to  de- 
ficiency in  the  salt  mixture.  Stepp  ascribed  the  limited  usefulness 
of  this  ration  to  deficiency  of  lipoids. 

Falta  and  Noeggerath  (1906)  attempted  to  maintain  adult 
white  rats  on  artificial  food  mixtures  and  inorganic  salts,  including 


858  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

calcium  phosphate.  Neither  the  rations  containing  phosphorus- 
free  proteins  nor  those  containing  organic  phosphorus,  as  of  casein, 
would  keep  the  animals  alive  indefinitely,  though  controls  fed  on 
milk,  milk  powder  or  horse  flesh  throve  without  incident. 

L.  Jacob  (1906)  kept  rats  as  long  as  124  days  on  artificial  mix- 
tures of  foodstuffs,  such  as  casein,  cane  sugar,  cellulose,  hog  fat  and 
salts.  Doves  fed  on  a  similar  mixture  did  not  do  well,  perhaps  be- 
cause the  feed  formed  a  soft  mass  in  the  crop. 

Knapp  (1908)  records  unsuccessful  attempts  to  maintain  rats 
on  artificial  foods  and  salts. 

Stepp  (1909)  sought  to  determine  the  office  of  the  lipoids  of 
bread,  on  the  growth  of  mice, » by  feeding  mice  on  bread  extracted 
with  alcohol  and  ether,  or  alcohol,  ether  and  chloroform,  in  compari- 
son with  others  which  received  the  residue  plus  the  extract.  The 
results  were  inconclusive ;  none  of  the  mice  throve. 

At  a  later  date  Stepp  (1911)  conducted  a  series  of  19  experi- 
ments on  the  growth  of  mice  on  various  diets  with  and  without  the 
presence  of  those  lipoid  bodies  which  are  soluble  in  alcohol  and 
ether.  Maintenance  of  life  and  live  weight  were  the  criteria  on 
which  conclusions  were  based.  Some  of  the  more  important  re- 
sults are  as  follows : 

Mice  die  when  fed  with  food  which  has  been  completely  freed 
from  lipoids.  The  length  of  life  is  increased  if  the  alcohol-ether 
soluble  constituents  are  added  to  the  extracted  food.  Fat  plays  no 
significant  role  itself,  though  there  are  in  butter  and  in  milk-fat 
very  small  quantities  of  important  alcohol-ether  soluble  materials. 
These  substances  are  present  in  greater  amounts  in  milk  plasma. 
Lecithin  and  cholesterin  seem  not  to  be  the  necessary  lipoids. 

That  unknown  food  constituents  may  possess  properties  of 
much  importance,  and  of  a  value  out  of  proportion  to  amount,  is  in- 
dicated by  experiments  by  Hopkins  (1912)  in  the  feeding  of  rats. 
When  added  to  rations  of  purified  products  such  as  pure  casein, 
starch,  cane  sugar,  lard  and  salts — a  very  small  amount  of  milk  had 
the  effect  to  maintain  normal  growth,  which  otherwise  soon  ceased. 
The  effect  was  shown  not  to  be  due  to  greater  amount,  energy  value 
or  palatability  of  the  ration  containing  the  small  amount  of  milk. 
The  nature  of  the  important  principle  was  not  determined. 

One  of  us  (E.  B.  F.)  has  had  similar  experience  in  feeding  ex- 
periments with  pigs  on  a  basal  ration  of  pearl  hominy,  wheat  gluten, 
blood  albumen  and  corn  bran,  to  which  were  added  various  pure 
phosphorus  compounds.  It  has  often  been  the  case,  with  rations 
other  than  those  containing  glycerophosphates,  that  in  order  to 
keep  the    pigs  alive  until    the  end    of  the  experiment    it  has  been 


PHOSPHORUS  METABOLISM  359 

necessary  to  introduce  a  small  amount  of  corn  into  the  ration.  The 
improvement  was  always  marked.  At  these  times  we  were  unable 
to  get  the  pigs  into  good  condition  by  the  addition  to  the  ration  of 
corn  oil,  mangel  wurzels,  potassium  citrate,  calcium  carbonate  or 
soda. 

Osborne  and  Mendel  (1911a)  in  their  first  year's  investigation 
on  feeding  with  isolated  food  substances  (reported  in  Carnegie  In- 
stitution Publication  156,  Part  I)  conducted  many  experiments  in 
the  feeding  of  white  rats  on  rations  of  pure  casein,  cane  sugar, 
starch,  lard,  agar  and  inorganic  salts.  Varying  results,  as  deter- 
mined by  the  salt  mixtures,  were  obtained.  Weight  was  main- 
tained and  nitrogen  stored  over  considerable  periods  of  time  with 
both  young  and  mature  rats,  but  without  significant  gain  in  weight, 
on  rations  containing  but  a  single  protein.  One  animal  was  main- 
tained for  217  days  on  a  diet  in  which  the  sole  protein  was  glutenin. 

In  the  second  year's  work,  as  reported  in  Part  II  of  the  above- 
mentioned  publication,  a  large  number  of  pure  proteins  were  com- 
pared, some  containing  phosphorus,  and  others  free  from  this  ele- 
ment. A  noteworthy  change  of  method  was  the  addition  to  the 
ration  of  "protein-free"  milk,  a  preparation  made  from  skim  milk  by 
precipitation  of  the  casein  with  hydrochloric  acid,  nitration,  and 
evaporation  of  the  whey. 

Active  growth  was  obtained  with  casein,  ovalbumin,  lactal- 
bumin,  edestin,  glutenin  and  glycinin,  each  as  the  sole  protein 
constituent  of  the  ration.  With  gliadin,  and  hordein  there  was 
little  or  no  growth,  and  with  zein  weight  was  not  maintained.  The 
use  of  protein-free  milk  rendered  this  series  of  experiments  the  first 
thoroughly  successful  attempt  to  induce  growth  in  rats  with  such 
isolated  food  substances.  The  maximum  normal  size  of  the  animals 
was  not  attained,  however,. as  the  authors  state  in  their  later  work 
on  fat-free  foods  (Osborne  and  Mendel,  1912b),  but  an  increase  to 
several  times  the  initial  weight  was  produced  on  rations  at  least 
essentially  free  from  purins  and  from  organic  phosphorus  com- 
pounds.    See  also  Osborne  and  Mendel,  1911b. 

In  the  article  entitled  "Ueber  Fiitterungsversuche  mit  isolierten 
Nahrungssubstanzen"  (1912b)  Osborne  and  Mendel  review  their 
previous  work  and  add  reports  of  further  similar  tests  of  the  ability 
of  rats  to  gain  in  weight  on  rations  in  which  certain  legume  and 
other  proteins  constituted  the  sole  source  of  protein  nutriment.  A 
table  is  presented  in  which  the  many  pure  proteins  tested  are 
arranged  in  the  order  of  the  gain  in  weight  induced  by  them,  in 
rats,  during  30  days.  In  the  order  named,  beginning  with 
those  which  cause  loss  in  weight  they  are  as  follows :  Phaseolin,  zein, 
gelatin  and  conglutin,  the  loss  in  weight  of  the  last-mentioned  being 


360  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

least.  Then  in  a  series  of  ascending  efficiency  are  mentioned  rye 
gliadin,  hordein,  pea  legumin,  wheat  gliadin,  vignin,  legumelin, 
hemp-seed  glutelin,  edestin,  glycinin,  wheat  glutenin,  cotton-seed 
globulin,  lactalbumin,  maize  glutelin,  excelsin,  squash-seed  globulin, 
ovalbumin,  casein,  and,  most  efficient  of  all,  ovovitellin. 

In  a  paper  on  "The  Role  of  Gliadin  in  Nutrition"  Osborne  and 
Mendel  (1912c)  showed  that  the  gliadins  of  wheat  and  rye,  as  well 
as  the  closely  related  hordein  of  barley,  suffice  for  the  maintenance 
of  rats  without  growth.  A  female  rat  after  178  days  with  gliadin 
as  its  sole  protein  gave  birth  to  four  young.  At  the  end  of  30  days 
3  were  removed.  The  one  remaining  with  the  mother,  however, 
failed  to  thrive  on  the  gliadin  food.  The  authors  consider  that  in 
this  experiment  there  must  have  been  a  synthesis  of  those  protein 
"building  stones"  which  were  lacking  in  the  intake  during  the  period 
of  gestation  and  lactation.  The  rat  only  maintained  its  weight 
previous  to  mating.  During  pregnancy  the  gain  in  weight  was  very 
rapid.  Subsequent  to  pregnancy,  however,  the  loss  in  weight  was 
pronounced,  and  the  rat  died  94  days  after  bearing  young,  her 
weight  at  that  time  being  only  a  little  more  than  half  that  at  the 
time  she  was  mated.      No  post-mortem  was  held. 

A  number  of  charts  are  presented  showing  the  history  of  ex- 
perimental animals  subsequent  to  the  period  of  normal  growth  on 
the  rations  containing  but  a  single  protein.  After  such  normal 
increase  in  weight,  as  previously  mentioned,  these  animals  reached  a 
period  of  rapid  decline  in  weight.  Body  analyses,  to  demonstrate 
that  the  normal  increase  in  weight  was  in  every  way  normal  in  char- 
acter, would  be  of  interest  in  this  connection. 

In  "Maintenance  Experiments  With  Isolated  Proteins"  these 
authors  state  that  "every  animal  has  sooner  or  later  declined  when 
fed  with  mixtures  of  isolated  and  purified  proteins,  carbohydrates 
and  fats  together  with  inorganic  matter  in  the  form  of  crystallized 
salts.  In  nearly  every  case  the  decline  has  been  sudden,  with 
strong  evidence  that  death  would  soon  have  ensued  had  not  the  food 
been  changed.  In  each  case  immediate  recovery  has  followed  a 
change  in  the  diet,  thus  showing  the  experimental  foods  to  be 
inadequate  for  prolonged  nutrition."  They  state,  however,  that 
"with  the  aid  of  'protein-free'  milk  it  is  possible  to  maintain  rats  for 
periods  equal  to  practically  their  entire  adult  lives  on  foods  contain- 
ing a  single  purified  protein."  They  also  note  the  work  of  Hopkins 
(1912)  who  shows  that  milk,  as  well  as  other  natural  food  materials, 
contains  unknown  substances  which,  even  in  very  small  quanti- 
ties, suffice  to  induce  normal  and  continued  growth,  for  several 
weeks,  at  least,  in  rats  maintained  on  artificial  mixtures  of  food  sub- 
stances which  are  otherwise  inadequate  for  growth. 


PHOSPHORUS  METABOLISM  361 

Twelve  rats  were  maintained  more  than  400  days,  and  5  rats 
more  than  500  days  on  rations  of  isolated  foodstuffs  free  from  more 
than  the  merest  possible  traces  of  purins  and  of  phosphoproteins. 

In  1912  Osborne  and  Mendel  published  a  series  of  four  papers  all 
bearing  on  the  subject  of  protein  synthesis,  and  incidentally  on  the 
synthesis  of  compounds  of  phosphorus.      The  method  of  experi- 
mentation was  modified  by  the  use  of  an  artificial  mixture  of  inor- 
ganic salts  and  milk  sugar  in  imitation  of  the  protein-free  milk  prep- 
aration which  had  made  possible  the  success  of  the  work  of  the 
previous  year.      This  artificial  preparation  proved  to  be  the  most 
efficient  salt  mixture  thus  far  compounded.      In  their    work    on 
"Feeding  Experiments  With  Fat-Free  Food  Mixtures"  (1912a)  they 
were  successful  in  causing  growth  of  young  white  rats  on  rations  of 
isolated  food  substances  which  were  entirely  free  from  fat,  and  at 
least  essentially  free  from  any  compounds  of  lipoid  nature,  the  pro- 
tein being  present  in  two  cases  as  casein,  in  another  as  casein  and 
edestin,  and  in  another  as  edestin  alone.    Phosphorus  was  present 
in  the  last-mentioned  only  in  the  authors'  "artificial  protein-free 
milk."      In  this  last  case  the  curve  of  growth  shows  that  the  rat  in- 
creased in  weight  to  five  times  the  initial  weight  in  60  days.  It  is 
difficult  to  believe  that  any  sort  of  redistribution  of  constituents 
could  make  possible  so  considerable  an  increase  in  body  weight  with- 
out extensive  synthesis  of  all  of  the  phosphorus-containing  com- 
pounds necessary  to  growth.      The  actual  proof  of  such  synthesis 
by  body  analysis  of  the  experimental  subjects  will  doubtless  be 
forthcoming  at  an  early  date.      That  the  growth  was  normal  in 
composition  was  not  in  any  way  proven. 

These  authors  were  unable,  however,  similarly  to  induce  growth 
in  mice  on  these  rations,  and  they  note  the  agreement  of  their 
experience  in  this  matter  with  that  of  Stepp. 

Osborne  and  Mendel  (1913)  once  more  report  their  failure  even 
to  maintain  mature  animals  indefinitely  on  artificial  foods  and  inor- 
ganic salts.  They  write  as  follows  (p.  132),  "It  is  true  that  in  sev- 
eral instances  we  have  succeeded  in  keeping  grown  rats  in  health  and 
in  apparent  nutritive  equilibrium  on  purely  artificial  food  mixtures 
over  periods  far  longer  than  the  experience  of  our  predecessors  had 
led  us  to  expect.  But  the  outcome  has  never  been  satisfactory  in 
the  sense  of  extending  over  what  may  be  considered  as  the  larger 
portion  of  the  life-span  of  an  adult  animal.  Successful  maintenance 
has  been  secured  only  when  the  animals  have  been  fed,  in  part  at 

least,  with  foods  containing  our  'protein-free  milk/ The 

superiority  of  the  latter  foods,  compared  with  any  purely  artificial 


362  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

food  mixture  in  repairing  the  depleted  body  weight  of  animals  that 
have  begun  to  decline  on  the  artificial  salt  mixtures  tested  is  beyond 
question." 

But  even  with  the  use  of  this  "protein-free  milk"  growth  may 
not  be  sustained  in  rats  in  a  normal  manner,  "a  few  stopping  after 
sixty  days  of  growth,  others  continuing  to  grow  for  100  days  or 
more.  After  normal  growth  stops,  the  animals  may  remain  in  con- 
stant weight  for  a  few  days,  or  grow  very  slowly,  and  then  suddenly 
decline  and  die  unless  a  change  is  made  in  the  diet The  con- 
clusion seems  inevitable,  therefore,  that  the  'protein-free  milk  foods' 
are  deficient  in,  or  completely  lack,  something  which  milk  contains 
and  which  is  indispensable  for  perfect  growth." 

The  importance  of  certain  inorganic  elements  not  usually  con- 
sidered in  nutrition  investigations,  namely  iodine,  manganese,  fluor- 
ine and  aluminium,  was  demonstrated  by  the  addition  of  very  small 
amounts  of  salts  of  these  elements  to  the  artificial  protein-free  milk 
previously  mentioned.  Much  better  results  in  feeding  experiments 
were  obtained  with  these  elements  present  than  in  their  absence, 
and  a  considerable  portion  of  the  favorable  results  obtained  with  the 
previous  use  of  artificial  protein-free  milk  must  be  considered  to 
have  been  dependent  on  inorganic  impurities  present  in  the  chemicals 
used  in  its  preparation. 

The  authors  show  that  butter  contains  the  principle  which  is 
present  in  milk,  lacking  in  their  "protein-free  milk,"  and  which  is 
essential  to  normal  growth. 

Hopkins  and  Neville  (1912)  also  fed  a  large  number  of  rats  on 
the  diet  used  by  Osborne  and  Mendel,  but  the  rats  ceased  to  grow 
before  the  attainment  of  full  normal  size. 

Ruth  Wheeler  (1913)  maintained  white  mice  in  health  for  6 
months  on  an  artificial  diet  containing  but  a  single  protein  (casein) . 

Stepp  (1913a)  experimented  on  mice,  with  various  lipoid-free 
diets,  plus  lipoids  of  diverse  sorts  and  conditions.  The  lipoids  of 
brain,  egg  yolk,  etc.,  possessed  life-saving  power  under  certain  con- 
ditions, but  this  capacity  is  lost  through  heating.  Stepp  speculates 
as  to  the  probability  of  the  essential  nutrient  being  lecithin.  Com- 
mercial lecithin,  however,  had  not  the  life-saving  power,  perhaps 
because  of  modification  in  the  course  of  separation  from  the  natural 
product  containing  it,  perhaps  because  another  phosphatid  is 
required. 

Stepp  (1913b)  has  studied  the  importance  to  animal  life  of  the 
lipoids,  phosphorized  and  otherwise,  as  separated  from  foods  by 
various  solvents.  Experimenting  with  mice — ether  seems  not 
to  remove  essential  nutrients  from  foodstuffs;  alcohol  ex- 
traction, however,  leaves  the    food    incapable    of    sustaining    life. 


PHOSPHORUS  METABOLISM  363 

Acetone  extract  of  egg  yolk  does  not  contain  the  essential  nutrients 
removed  by  alcohol,  nor  does  an  alcohol  extract  of  the  acetone 
extraction  residue.  Thus  it  appears  that  acetone  removes  a  part, 
but  not  all,  of  the  essential  nutrients  of  egg  yolk  which  are  soluble 
in  alcohol.  A  mixture  of  lecithin,  cholesterol,  cephalin,  cerebron  and 
phytin  can  not  compensate  for  the  substances  extracted  from  the 
diet  by  alcohol-ether. 

McCollum  and  Davis  (1913a)  obtained  normal  gain  in  weight  of 
rats  during  75-100  days  on  a  ration  of  casein,  dextrin,  agar-agar 
and  inorganic  salt  mixtures  similar  to  the  mineral  content  of  eggs 
or  milk;  Such  mixtures,  however,  do  not  support  growth  indef- 
initely. In  a  later  paper  (1913b)  these  authors  state  that  beyond 
100-120  days  little  or  no  increase  in  weight  can  be  produced  on  such 
rations  as  the  above.  That  animals  fed  on  these  rations  of  puri- 
fied food  substances  are  in  a  physiological  state  which  is  nearly  nor- 
mal was  concluded  from  the  fact  that  three  female  rats  produced 
young  after  being  fed  only  casein,  carbohydrates,  lard  and  salt  mix- 
tures for  periods  of  108,  127  and  142  days,  respectively.  These  rats 
had  made  approximately  normal  growth  for  about  80  days  on  this 
ration.  In  none  of  these  cases,  however,  were  the  mothers  able  nor- 
mally to  nourish  the  young.  The  authors  believe  that  cessation  of 
growth  on  such  rations  as  the  above  is  due  to  the  animals'  "running 
out"  of  some  essential  organic  complex,  which  they  determine  to  be 
present  in  the  ether-extract  of  egg  and  of  butter.  Lard  and  olive  oil 
were  not  similarly  efficacious.  The  feeding  of  lecithin  and  choles- 
terin  gave  results  which  the  authors  characterize  as  "very  doubt- 
ful."     The  data  were  not  presented. 

In  applying  to  human  nutrition  and  to  live-stock  feeding  the 
results  of  these  experiments  with  rats  and  i  mice,  it  is  our  feeling 
that,  in  spite  of  the  fundamental  similarity  of  the  nutritive  process- 
es of  all  vertebrates,  we  should  consider  the  possibility  of  important 
differences  in  the  metabolism  of  remotely  related  forms. 

In  the  literature  on  this  subject  we  note  a  marked  tendency  on 
the  part  of  some  whose  experience  with  animals  has  been  confined 
to  laboratory  work  with  a  very  few  species,  to  ascribe  to  animals 
generally,  without  qualification,  the  physiological  capacities  of 
each  species.  Many  important  differences  are  known  to  exist  in  the 
metabolism  of  vertebrates.  As  a  single  instance  of  this — some 
mice  can  live  without  drinking.  While  consuming  only  air-dry 
foods  the  metabolic  water  produced  by  oxidation  within  the  tissues 
is  adequate  for  the  performance  of  all  necessary  functions.  Such 
a  fact  should  convince  us  of  the  need  of  conservatism  in  the  practi- 


364  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

cal  application  of  some  of  our  experimental  results  before  they  have 
been  shown  actually  to  possess  significance  in  relation  to  the  animal 
and  the  conditions  of  interest. 

SUMMARY  ON  NUTRITIVE  VALUES  OF  ORGANIC  AND  INORGANIC 

PHOSPHORUS 

In  the  light  of  our  present  understanding,  we  must  deplore  the 
imperfect  and  inconclusive  character  of  much  of  the  work  on  this 
subject.  That  the  evidence  is  not  more  satisfactory  is  because  it 
has  been  only  with  the  gradual  accumulation  of  knowledge  in  this 
field  that  investigators  have  come  to  realize  the  difficulties  and  the 
requirements  of  a  positive  solution  of  this  problem. 

Considering  the  evidence  as  a  whole,  however,  there  is  much  of 
value,  and  even  the  more  imperfect  work,  in  the  aggregate,  produces 
a  composite  impression  which  can  hardly  fail  to  affect  opinion ;  and 
that  this  should  be  true  is  perfectly  proper,  unless  there  be,  through- 
out the  work,  a  neglect  of  absolutely  essential  considerations.  Can- 
dor requires  that  we  admit  one  such  possibility.  In  many  experi- 
ments, at  least,  there  has  been  no  guarantee  of  the  purity  of  the 
organic  compounds  used,  and  no  particular  effort  to  guard  against 
the  influence  of  other  useful  compounds  associated  with  the  organic 
phosphorus  in  the  natural  products  and  also  in  the  isolated  com- 
pounds as  used  in  these  investigations. 

This  consideration  we  must  bear  in  mind.  It  does  not  invalid- 
ate all  of  the  work  on  this  subject,  but  should  serve  to  increase  the 
scientific  conservatism  with  which  we  naturally  regard  the  evidence. 
Considering  that  we  have  sufficiently  qualified  our  conclusions  we 
shall  point  out  the  apparent  significance  of  the  foregoing  investiga- 
tions. 

One  point,  at  least,  in  this  connection,  we  may  regard  as  defin- 
itely established.  Considering  the  phosphorus  requirement  as  a 
whole,  an  amount  of  organic  phosphorus  equal  to  a  very  small  part 
of  the  total  is  sufficient  for  growth  and  reproduction,  provided  inor- 
ganic phosphorus  be  present  in  sufficient  amount.  This  fact  is 
not  likely  to  be  questioned,  in  consideration  of  the  very  small  propor- 
tion of  the  total  phosphorus  of  the  animal  which  is  present  in  organ- 
ic combination. 

That  organic  phosphorus  is  absolutely  essential  to  any  animal 
has  not  been  demonstrated.  The  proof  that  inorganic  phosphorus 
can  serve  all  of  the  purposes  for  which  any  animal  needs  phosphorus 
is  incomplete.  There  is  much  evidence  to  imply  that  with  some 
species,  at  least,  some  organic  phosphorus  compounds  are  more 
useful  than  is  inorganic  phosphorus  in  the  sense  of  being  more 
readily  and  economically  utilized,  and  of  maintaining  a  higher  state 


PHOSPHORUS  METABOLISM  365 

of  vitality  as  revealed  by  tissue  enzyme  estimations,  the  difference 
probably  depending,  in  part  at  least,  on  the  fact  of  the  partial 
absorption  and  utilization  of  organic  phosphorus  compounds  as  such, 
without  complete  digestive  cleavage. 

An  important  aspect  of  this  problem  is  afforded  by  the  results 
of  feeding  experiments  on  rats  and  mice,  with  simple  purified 
nutrients,  and  by  the  results  of  investigations  of  the  cause  of  beri- 
beri (see  Beriberi).  In  the  light  of  these  studies  the  synthesis 
of  organic  phosphorus  compounds  in  a  normal  manner  seems  to  be 
dependent  on  the  presence  of  minute  quantities  of  certain  unknown 
compounds  which  are  found  in  natural  foodstuffs,  and  which  act  like 
enzymes,  catalyzers  or  activators.  Such  essential  compounds  are 
found  in  milk  and  in  eggs  and  other  natural  foods  of  both  plant  and 
animal  origin.  Lack  of  such  nutrient  principles  in  white  rice  and 
patent  flour  has  been  shown  to  be  the  cause  of  beriberi,  and  Funk 
suggests  that  a  deficiency  of  the  food  in  these  unknown  compounds, 
for  which  he  proposes  the  name  "vitamines,"  produces  a  predisposi- 
tion to  many  other  diseases  such  as  polyneuritis  in  fowls,  epidemic 
dropsy,  experimental  scurvy  in  animals,  infantile  scurvy,  ship  beri- 
beri and  pellagra.  From  the  facts  of  the  occurrence  of  these  sub- 
stances in  rice  polish,  and  their  absence  from  polished  rice ;  and  also 
their  presence  in  whole  wheat  flour,  and  absence  from  white  flour, 
we  must  conclude  that  in  vegetable  foods  their  distribution  is 
localized  in  certain  tissues. 

It  therefore  seems  not  at  all  unlikely  that  the  many  demonstra- 
tions of  superior  nutritive  value  of  organic  to  inorganic  phosphorus 
compounds  have  been  influenced  by  other  beneficial  substances 
occurring  in  association  with  them  in  natural  foods,  and  contained 
as  impurities  in  these  organic  phosphorus  compounds  as  isolated  and 
used  in  nutrition  investigations.  As  to  the  relative  importance  of 
this  factor  and  others  we  are  as  yet  unprepared  to  make  positive 
assertions ;  but  these  recent  studies  at  least  raise  the  question  as  to 
whether  the  apparent  superiority  of  organic  to  inorganic  phospho- 
rus compounds  is  due  to  these  organic  compounds  by  themselves,  or 
whether  their  superiority  is  dependent  on  minute  quantities  of  cer- 
tain associated  compounds.  However  this  question  may  be  settled, 
these  studies  certainly  suggest  that,  if  the  natural  organic  phos- 
phorus compounds  are  not  of  superior  usefulness,  or  are  not  essen- 
tial to  the  maintenance  of  growth  in  animals,  then  other  nutrients 
associated  with  them  in  the  natural  foods  are  essential,  and  the 
result,  therefore,  is  to  put  a  new  emphasis  on  the  value  of  the 
natural  organic  foodstuffs  as  compared  with  inorganic  or  artificially 
synthesized  nutrients  and  certain  manufactured  foods. 


366 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


SOME    COMMON    FOODS    IN    RELATION    TO    PHOSPHORUS 

METABOLISM 

MISCELLANEOUS  EXPERIMENTS  WITH  MEN  AND  ANIMALS 

The  relative  capacities  of  common  foods  to  supply  the  phospho- 
rus requirements  of  animals  is  a  subject  of  much  practical  import- 
ance on  which  we  have  comparatively  little  evidence,  and,  in  view  of 
the  fact  that  the  comparisons  must  be  made,  if  they  are  to  be  of 
practical  value,  not  between  individual  foodstuffs,  but  between 
mixed  rations,  wholly  satisfactory  evidence  in  this  field  is  not  to  be 
expected.  However,  very  imperfect  evidence  is  not  necessarily 
without  value,  and  is  often  the  only  sort  obtainable  with  reference 
to  our  most  important  problems. 

EXPERIMENTS  WITH  HUMAN  BEINGS 

Surmont  and  Dehon  (1903)  conducted  24-hour  balance  experi- 
ments with  human  subjects  in  the  comparison  of  white  bread  and 
whole  wheat  bread,  eaten  with  distilled  water  only.  A  portion  of  the 
results  are  as  follows : 
NITROGEN    AND    PHOSPHORUS    BALANCES    COMPARING    WHOLE 

WHEAT  BREAD  WITH  WHITE  BREAD,   WITH  TWO   HUMAN 
SUBJECTS  IN  TWENTY-FOUR  HOUR  EXPERIMENTS— Grams 


Bread 
eaten 

Nitrogen 

Phosphorus  (P2O5) 

bread 

Food 

Urine 

Feces 

Balance 

Food 

Urine 

Feces 

Balance 

White 

Whole  wheat. 

Whole  wheat. 

680 
605 

950 
950 

8.357 
11.942 

11.941 
18.230 

7.089 
11.445 

12.130 
12.440 

1.573 

2.485 

3.835 
7.021 

-0.305 

-0.988(1) 

-4.024 
—1.231 

1.455 
6.419 

1.990 
9.471 

1.859 
5.265 

1.517 
6.422 

0.930 
2.683 

1.118 
3.657 

-1.334 
-1.529 

-0.645 
-0.608 

(*)      Should   be  — 1.988   to   correspond  with  other  data. 

The  authors  state  that  the  two  lots  of  white  bread  contained 
1.229  percent  and  1.257  percent  nitrogen,  and  0.214  and  0.199  per- 
cent P,05 ;  the  whole  wheat  bread  1.974  and  1.919  percent  nitrogen, 
and  1.061  and  0.997  percent  P205- 

A  larger  proportion  of  the  phosphorus  of  the  white  bread  was 
found  in  the  feces  than  of  the  whole  wheat  bread.  The  apparent 
digestibility  of  the  nitrogen  was  about  the  same  in  both  cases.  The 
periods,  however,  were  so  short  that  the  results  have  only  corrobor- 
ative value.  The  very  considerable  phosphorus  deficit  on  the 
whole  wheat  bread  diet,  with  an  intake  of  6.4  gm.  P205  in 
one  case  and  9.47  gm.  in  another  is  remarkable,  since  these  amounts 
of  phosphorus  are  much  more  than  enough  to  maintain  phosphorus 
equilibrium  under  ordinary  conditions.  The  very  short  periods  give 
results  indicating  habit  of  phosphorus  metabolism  rather  than 
phosphorus  requirements. 


PHOSPHORUS  METABOLISM 


367 


Surmont  and  Dehon  (1904)  continued  their  study  of  white  and 
whole  wheat  bread  at  a  later  date.  A  part  of  their  figures  are  as 
follows : 

AVERAGE    DAILY  NITROGEN   AND  PHOSPHORUS    BALANCES   WITH 

TWO  MEN  COMPARING  WHITE  BREAD  WITH  WHOLE  WHEAT 

BREAD  IN  THREE-DAY  PERIODS— Grams 


Kind  of  bread 

Nitrogen 

Phosphorus  (P2O5) 

Food 

Urine 

Feces 

Balance 

Food 

Urine 

Feces 

Balance 

12.213 
U.718 

12.213 
11.718 

10.377 
10.227 

12.034 
11.976 

2.458 
2.566 

2.414 
2.527 

-0.622 
-1.076 

-2.235 

-2.785 

6.384 
8.163 

6.384 
8.163 

4.004 
5.081 

4.082 
5.125 

2.311 

2.887 

2.238 
2.870 

+0.069 
+0.195 

+0.066 
+0.168 

The  subjects  were  first  brought  to  nitrogen  equilibrium  on  a 
constant  mixed  diet ;  then  followed  the  3-day  experiments  on  whole 
wheat  bread  as  a  part  of  the  mixed  diet.  Nitrogen  equilibrium  was 
again  established  in  an  intermediate  period,  and  then  followed  the 
period  in  which  white  bread  was  consumed  with  a  mixed  diet. 

The  authors  concluded  that  the  phosphorus  of  whole  wheat 
bread  is  as  well  absorbed  as  that  of  white  bread,  and  that  the 
coefficient  of  retention  of  the  phosphorus  of  whole  wheat  bread  is 
higher  than  that  of  the  phosphorus  of  white  bread.  The  assimi- 
lation of  the  nitrogen  of  the  whole  wheat  bread  was  found  somewhat 
lower  than  with  the  white  bread. 

Newman,  Robinson,  Hainan  and  Neville  (1912)  compared  white 
and  whole  wheat  breads  in  digestion  experiments  of  7  days'  dura- 
tion with  four  men.  The  phosphorus  of  the  whole  wheat  bread, 
being  present  in  much  greater  quantity  than  in  the  white  bread,  was 
apparently  absorbed  as  efficiently,  and  therefore  in  larger  proportion 
to  absorbed  nitrogen,  thus  approaching  the  ratio  of  these  constitu- 
ents as  absorbed  from  efficient  mixed  dietaries.  The  nitrogen  of 
the  white  bread  was  considered  to  be  absorbed  3.5  percent  more 
efficiently  than  that  from  the  whole  wheat  bread. 

Schlossmann  and  Moro  (1904)  attempted  to  compare  metabo- 
lism of  a  28-year-old  man  on  a  diet  of  cow's  milk  with  the  same  on  a 
diet  of  human  milk,  but  psychic  factors  rendered  results  patho- 
logical on  the  latter  diet,  and  the  figures  therefore  have  no 
especial  value.  In  a  2-day  period  on  cow's  milk,  cream,  milk  sugar  and 
cognac,  with  a  daily  intake  of  4.487  gm.  CaO  and  6.113  gm.  P205, 
there  was  a  storage  of  1.60  gm.  CaO  and  1.33  gm.  P205.  The  sub- 
ject weighed  59.1  kg. 


368  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Wiley,  and  associates  (1904,  1906,  1907,  1908a,b)  studied 
metabolism  in  healthy  young  men  as  influenced  by  various  food 
preservatives.  Their  results  on  phosphorus  metabolism  were  rather 
variable,  and  were  not  striking. 

From  the  administration  of  3  gm.  boric  acid,  or  equivalent 
amounts  of  borax  daily,  it  would  appear  that  this  preservative 
slightly  decreases  phosphorus  retention. 

Salicylic  acid  and  salicylates  in  quantities  of  0.210  gm. — 2.00 
gm.  per  day  appeared  to  increase  to  a  slight  extent  the  absorption  of 
phosphorus  from  the  intestine,  and  to  lead  to  its  retention  in  the 
body.  The  positive  phosphorus  balance  was  somewhat  greater  in 
amount  during  the  preservative  period  than  either  before  or  after. 

Sulphurous  acid  and  sulphites  in  quantities  of  0.171  gm.  to  1.020 
gm.  (S02)  per  day  increased  the  feces  phosphorus.  The  authors 
assume  that  this  increase  is  due  to  inhibited  absorption  from  the 
intestine. 

Benzoic  acid  and  sodium  benzoate  in  quantities  of  1.0 — 2.5  gm. 
daily  of  benzoic  acid,  or  equivalent  amounts  of  sodium  benzoate,  also 
caused  a  slight  increase  in  feces  phosphorus ;  while  formaldehyde  in 
quantities  of  100-200  mg.  daily  caused  a  slight  increase  in  phospho- 
rus excretion,  with  a  considerable  deflection  of  phosphorus  from 
feces  to  urine. 

The  experimental  periods  were  of  considerable  duration  and  the 
scope  of  the  work  extensive.  The  feces  were  not  marked  at  the 
beginning  and  end  of  periods. 

Slowtzoff  (1909a,  1909b)  reports  a  comparison  of  calcium,  mag- 
nesium and  phosphorus  metabolism  on  a  diet  containing  meat  with 
the  same  after  the  substitution  of  fish  for  meat.  The  experiment 
covered  two  periods  of  four  days  each.  Under  the  influence  of  the 
fish  the  phosphorus  and  magnesium  retention  was  increased,  while 
the  calcium  retention  was  decreased.  The  author  considers  especially 
characteristic  the  increase  of  the  calcium  (about  60  percent)  and 
magnesium  (about  44  percent)  of  the  urine  above  the  amount 
present  during  the  meat  period.  Of  this  article  we  have  seen 
abstracts  only. 

The  most  important  demonstration  of  specific  effects  of  com- 
mon foods  on  phosphorus  metabolism  is  certainly  the  recent  discov- 
ery of  the  cause  of  beriberi.  In  this  disease  profound  disorganiza- 
tion of  the  nervous  system  is  due  to  the  lack  of  certain  important 
activating  principles  in  polished  rice.  For  a  discussion  of  this 
matter  see  Beriberi. 


PHOSPHORUS  METABOLISM  369 

EXPERIMENTS  WITH  DOMESTIC  ANIMAIiS 

Verdeil  (1849)  analyzed  the  blood  ash  of  different  species  of 
animals,  and  of  dogs  on  various  rations.  He  concluded  that  a  differ- 
ence of  feed  shows  quickly  in  the  composition  of  the  blood,  and  that 
in  the  blood  of  animals  eating  meat,  bread  or  grains  alkali  phos- 
phates are  abundant,  and  carbonates  almost  lacking,  while  with 
herbivora  the  opposite  relation  holds. 

Landsteiner  (1892)  found  no  significant  differences  in  the  blood 
ash  of  rabbits  fed  on  milk,  and  those  fed  on  hay  for  2>y%  months. 

According  to  Hart,  McCollum  and  Fuller  (1909),  the  blood  of 
four  pigs  that  had  received  different  amounts  and  kinds  of  phospho- 
rus compounds  in  the  diet  showed  a  phosphorus  range  from  0.24  to 
0.33  percent  P  (aver.  0.29  percent)  of  the  air-dry  matter.  The  cal- 
cium ranged  from  0.026  to  0.038  percent  Ca,  with  an  average  of 
0.033  percent. 

In  the  work  of  E.  B.  Forbes  and  associates  (1914)  the  minerals 
(including  phosphorus)  of  the  blood  were  found  to  vary,  in  marked 
and  consistent  ways  in  accord  with  the  character  of  the  food.  Ash 
varied  between  0.90  and  1.04  percent,  phosphorus  between  0.38  and 
0.64  percent,  inorganic  phosphorus  between  0.009  and  0.019  percent, 
calcium  between  0.0061  and  0.0085  percent,  magnesium  between 
0.0034  and  0.0059  percent,  potassium  between  0.172  and  0.230  per- 
cent, sodium  between  0.19  and  0.29  percent,  sulphur  between  0.120 
and  0.166  percent  and  chlorine  between  0.24  and  0.27  percent. 

Alquier  (1905-6),  from  feeding  experiments  on  horses,  conclud- 
ed that  beet  molasses  would  increase  the  solubility,  and  therefore 
the  assimilability,  of  the  phosphorus  of  the  ration,  through  the 
action  of  the  considerable  amounts  of  sodium  and  potassium  present 
in  this  food.  On  the  assumption  that  the  greatly  increased  amounts 
of  sodium  and  potassium  introduced  into  the  ration  by  the  molasses 
replaced,  in  a  measure,  the  calcium  of  calcium  phosphate,  it  seems 
quite  possible  that  the  suggestion  is  in  accord  with  the  facts. 

E.  B.  Forbes  (1909)  studied,  in  three  experiments  involving  120 
pigs,  the  specific  effects  of  a  number  of  common  foodstuffs,  especial- 
ly corn  (maize) ,  on  the  bodies  of  pigs,  largely  as  to  relations  of  fat, 
water  and  phosphorus.  Results  were  obtained  from  the  gain  in 
live  weight,  and  in  weight  of  parts  and  organs,  analyses  of  the 
tenderloin  (psoas  magnus)  muscles,  livers,  kidneys,  and  cross  sections 
of  the  body  at  the  6th  rib,  and  also  of  the  foodstuffs  used. 
The  first  experiment  involved  65  pigs  in  13  lots  of  5  each.  The 
feeding  lasted  60  days,  and  the  pigs  in  the  several  lots  gained  in 


370  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

weight  from  one  pound  to  1.7  pounds  per  head  and  day.  This  experi- 
ment was  conducted  in  two  series  of  6  lots,  one  receiving  food  ad 
libitum,  and  the  other  receiving  restricted  amounts  of  the  same 
foods,  one  lot  being  killed  as  a  control  at  the  beginning  of  the  experi- 
ment. Two  lots  (one  in  each  series)  received  corn  alone,  and  of  the 
remaining  ten  lots,  two  each  received  corn  supplemented  by  linseed 
oil  meal,  wheat  middlings,  corn  germ  .  oil  meal,  soy  beans  and 
digester  tankage. 

The  muscular  increase  was  generally  in  the  order  of  the  phos- 
phorus content  of  the  rations,  except  with  the  lots  which  received 
tankage,  the  phosphorus  of  which  was  present  mostly  as  bone. 

The  gain  of  fat  and  muscle  were  reciprocal;  that  is, •  arranging 
the  lots  according  to  gain  in  fat  arranged  them  inversely  as  the  gain 
in  muscle.  These  facts  suggest  that  common  foods  do  not  contain 
as  much  phosphorus  as  is  essential  to  maximum  growth  of  muscular 
tissue  and  skeleton.  Lecithin  phosphorus  seemed  not  to  be  the 
dominant  factor  in  this  consideration. 

Corn,  which  is  characterized  by  low  protein,  calcium  and  phos- 
phorus contents,  in  comparison  with  better-balanced  rations,  pro- 
duced under-sized,  over-fat  animals,  with  small  viscera,  and  deficient 
muscular  development,  and  bones  which  lacked  in  size,  strength 
and  ash  per  unit  of  volume.  The  muscles  were  characterized  by  low 
protein  and  low  ash  content,  and  high  content  of  f at^  while  the  water 
in  the  tissue  as  a  whole  was  low,  but  in  the  fat-free  meat  was  high. 
These  effects  are  due  especially  to  lack  of  protein  and  of  calcium, 
which  seem  to  limit  the  usefulness  of  the  phosphorus. 

The  second  and  third  experiments  involved  55  animals  in  12  lots, 
commonly  of  5  each.  They  were  conducted  in  the  same  general 
way  as  the  first,  but  the  rations  were  composed  of  pearl  hominy  and 
blood  flour  to  which  were  added  certain  phosphorus-containing 
preparations  of  interest.  These  were  (1)  water  extract  of  wheat 
bran,  containing  an  abundance  of  inosite-phosphoric  acid,  (2)  bone 
flour,  (3)  lecithin  and  (4)  sodium  phosphate.  The  same  amount  of 
phosphorus  was  fed  in  disodium  phosphate  as  in  lecithin.  In  these 
later  experiments  studies  were  also  made  on  the  growth  and  compo- 
sition of  the  bones  (humeri).  For  numerical  data  see  tables  on 
pages  371  to  374. 

The  water  extract  of  wheat  bran  appeared  to  contribute  to  the 
muscle-producing  capacity  of  the  ration,  and  bone  flour  was  decided- 
ly less  useful  in  this  way. 

Marked  characteristics  of  the  muscular  tissue  produced  in  these 
experiments  were  the  low  phosphorus  content  (0.195-0.252  percent) 
of  the  muscle  produced  by  rations  containing  the  bran  extract,  and 


PHOSPHORUS  METABOLISM 


371 


the  very  high  phosphorus  content  (0.352  percent)  of  the  muscle 
produced  by  the  ration  containing  lecithin. 

In  Experiment  II  the  bones  of  the  pigs  which  had  received  bone 
meal  exceeded  all  others  in  volume,  total  ash,  ash  per  c.c.  of  volume, 
and  breaking  strength.  Clearly  the  bone  meal  was  of  direct  value 
as  a  source  of  nutriment.  Observation  determined  the  fact  that 
the  deposit  of  bone  salts  was  largely  at  the  inside  of  the  walls  of 
the  bones.  In  some  cases  this  interior  thickening  of  the  walls 
proceeded  almost  to  the  obliteration  of  the  marrow  space.  Certain- 
ly there  is  a  much  less  definite  upper  limit  for  the  mineral  content  of 
the  bones  than  for  other  tissues.  This  is  in  harmony  with  their 
prominent  storage  function  for  mineral  nutriment. 

FOODS  AND  MINERAL  NUTRIENTS  CONSUMED  (Forbes,  1909) 

EXPERIMENT  H 


EXPERIMENT  HI 


Lot 

Rations 
Grams 

Cal- 
cium 

Grams 

Mag- 
nesium 

Grams 

Potas- 
sium 

Grams 

Sodium 
Grams 

Sul- 
phur 

Grams 

Phos- 
phorus 

Grams 

Chlo- 
rine 

Grams 

Excess 

normal 

acid 

C.  C. 

1 

Com ,.  2114.2 

.189 

2.326 

5.920 

.973 

2.981 

6.237 

.846 

217.90 

2 

Hominy. 1991.8 

Blood-flour 165.6 

1.092 

3.500 

7.239 

1.358 

4.236 

6.641 

1.491 

149.11 

Bran-extract,  (Larger  amt.)  2039.4 

3 

Hominy. 2068.9 

Blood-flour 187.3 

Lecithin 4.00 

.822 

.504 

2.166 

.921 

3.728 

1.562 

1.518 

198.50 

4 

Hominy. 2074.3 

.936 

1.675 

4.140 

1.100 

3.980 

3.429 

1.531 

Blood-flour 181.4 

176.50 

Bran-extract,  ( smaller  amt. )  779 . 3 

5 

Hominy. 1821.2 

Blood-flour 162.4 

5.747 

.552 

1.920 

.915 

3.286 

3.610 

1.337 

54.91 

6 

Hominy 2068.4 

.822 

.504 

2.166 

1.152 

3.728 

1.562 

1.518 

188.48 

; 

1426.9 

.134 

1.571 

3.998 

.657 

2.013 

4.212 

.571 

147  14 

3 

Blood-flour 

1548.6 

171.9 

.842 

1.272 

3.144 

.958 

3.214 

2.655 

1.245 

138  63 

.......    589.7 

4 

Blood-flour 

1570.4 

174.6 

3.406 

.463 

1.684 

.579 

3.054 

2.390 

1.247 

:  ] .  gfj 

11.0 

5 

1286.9 

143.3 

.616 

.332 

1.378 

.672 

2.500 

0.934 

1.021 

122.70 

372 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


In  Experiment  III  the  pigs  were  younger  than  in  Experiment  II, 
and  could  be  persuaded  to  eat  but  a  very  small  amount  of  the  bone 
meal,  the  pigs  on  the  ration  of  corn  alone  receiving  more  phosphorus 
than  the  lot  on  the  basal  ration  plus  bone  meal.  The  bone  data  from 
this  experiment,  therefore,  were  less  significant. 

The  pigs  receiving  the  maximum  amount  of  bran  extract  that 
they  would  tolerate,  and  receiving  in  this  ration  nearly  twice  as 
much  phosphorus  as  another  lot  which  was  fed  less  of  the  extract, 
had  smaller  muscles  and  also  smaller  bones,  the  bones  containing 
slightly  less  ash  per  cubic  centimeter  of  volume,  and  the  muscles 
less  phosphorus  than  in  the  lot  which  received  less  of  this  supple- 
ment. The  bran  extract  apparently  contained  some  constituent 
which,  when  present  in  excess,  tended  to  restrict  the  utilization  of 
the  phosphorus  of  the  ration.  This  extract  was  very  rich  in  mag- 
nesium. 

ANALYSES  OF  TENDERLOIN  MUSCLES  (Forbes,  1909)— Percent 

EXPERIMENT  II 


Lot 


Rations 


Water 


Pratein 


Fat 


Ash 


Phos- 
phorus 


Propor- 
tion of 
phos- 
phorus 

to 
protein 


Phos- 
phorus 
in  ash 


Corn. 


Hominy;  blood-flour;  bran-extract, 
(larger  amount) , 


Hominy;  blood-flour;  lecithin 

Hominy;  blood  flour;  bran-extract, 
(smaller  amount) 


Hominy;  blood-flour;  bone-meal.. . 
Hominy;  blood-flour;  sodium  phosphate 
Check  lot 


73.51 

73.62 
74.63 

72.74 
72.83 

72.87 
73.57 


19.73 

18.85 
15.78 

20.67 
20.13 
20.74 
18.53 


5.17 

5.17 
4.27 

4.22 
5.12 
5.14 
4.97 


1.09 

1.13 
1.10 

1.21 
1.08 
1.10 
1.10 


.264 

.195 
.352 

.252 
.233 
.264 
.247 


1.34 

1.04 
2.23 

1.22 
1.16 
1.27 
1.33 


24.22 

17.35 
32.00 

20.83 
21.57 
24.00 
22.45 


EXPERIMENT  III 


1 

73.39 
76.35 
73.74 
73.20 
72.81 

17.68 
19.42 
21.85 
18.30 
20.39 

6.16 
3.04 
3.90 
4.46 
4.52 

1.07 
1.13 
1.03 
1.12 
1.15 

.228 
.238 
.195 
.222 
.228 

1.29 
1.23 
.81 
1.21 
1.12 

21.31 

2 

Check  lot 

21.06 

3 
4 
5 

Hominy;  blood-flour;  bran-extract  .... 
Hominy;  blood-flour;  bone-flour 

18.93 
19.82 
19.83 

This  series  of  experiments  demonstrates  a  susceptibility  of  the 
animal  body  to  change  of  composition  as  a  result  of  the  character  of 
the  food  which  is  often  underestimated  and  still  more  frequently 
denied.  As  an  instance  of  this  fact  the  above  and  the  following 
tables  are  submitted. 


PHOSPHOEUS  METABOLISM 


373 


From  these  tables  we  note  that  the  phosphorus  in  muscle  varied 
in  amount,  expressed  as  percent  of  protein,  from  1.04  to  2.23  per- 
cent, and  expressed  as  percent  of  ash,  from  17.35  to  32  percent.  In 
liver  the  phosphorus  varied  between  1.57  and  2.10  percent  of  the 
protein,  and  between  24.18  and  28.23  percent  of  the  ash. 

In  the  kidneys,  the  phosphorus  varied  between  1.32  and  1.99 
percent  of  the  protein,  and  between  18.47  and  27.90  percent  of  the 
ash. 

These  variations  in  gross  proximate  analysis  are  certainly 
sufficient  to  indicate  profound  changes  in  functional  efficiency. 
Enzyme  estimations  in  connection  with  such  studies  would  probably 
be  of  value. 

ANALYSES  OF  LIVERS  (Forbes,  1909)— Percent 

EXPERIMENT  II 


Lots 


Rations 


Corn „ 

Hominy;  blood-flour;  bran-extract, 
(larger  amount) 


Hominy;  blood-flour;  lecithin 

Hominy;  blood-flour;  bran-extract, 
(smaller  amount) 


Hominy;  blood-flour;  bone-meal.. 
Hominy;  blood-flour;  sodium  phosphate 
Check  lot 


Water 

Protein 

Pat 

Ash 

Phos- 
phorus 

Propor- 
tion of 
phos- 
phorus 

to 
protein 

74.24 

18.83 

2.81 

1.27 

.320 

1.70 

73.52 

17.19 

2.46 

1.37 

.356 

2.07 

71.98 

18.72 

2.90 

1.29 

.364 

1.94 

71.96 

20.13 

3.01 

1.32 

.364 

1.81 

71.81 

19.45 

2.65 

1.39 

.361 

1.85 

72.05 

20.58 

2.85 

1.34 

.324 

1.57 

71.48 

14.61 

1.84 

1.17 

.291 

1.99 

Phos- 
phorus 
in  ash 


25.20 

**.  I 
26.00 
28.22 

27.58 
25.97 
24.18 
24.87 


EXPERIMENT  III 


Corn 

Check  lot 

Hominy;  blood-flour;  bran-extract. 
Hominy;  blood-flour;  bone-flour.. . . 
Hominy;  blood-flour 


70.77 

16.08 

2.32 

1.21 

.338 

2.10 

71.21 

18.12 

3.01 

1.33 

.338 

1.87 

71.65 

20.44 

2.49 

1.30 

.367 

1.80 

70.43 

19.31 

2.03 

1.30 

.366 

1.90 

70.64 

20.97 

2.26 

1.34' 

.346 

1.65 

27.93 
25.41 
28.23 
28.15 
25.82 


See  also  the  tables  on  the  following  page. 

Schkarin  (1910)  studied  the  influence  of  the  diet  of  mother 
dogs  on  the  development  of  suckling  pups.  No  characteristic  effects 
were  observable  of  the  diets  of  (1)  vegetable  food,  (2)  meat  and 
milk,  and  (3)  eggs  and  milk.  The  reason  for  negative  results  lies 
in  the  impossibility  of  modifying  the  composition  of  the  milk,  to  any 
considerable  extent,  through  the  character  of  the  food. 

Fingerling  (1911a)  attempted  to  solve  the  question  of  the  bet- 
ter utilization  by  ruminants  of  the  phosphorus  compounds  of  cereals 
and  mill  feeds  than  of  roughage.  After  various  tests,  which  threw 
no  light  on  the  matter,  he  fed,  in  comparison,  fresh  grass  and  hay 


374 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


ANALYSES  OF  KIDNEYS  (Forbes,  1909)— Percent 

EXPERIMENT  II 


Lots 


Rations 


Corn i 

Hominy;  blood-flour;  bran-extract, 
(larger  amount) , 


Hominy;  blood-flour;  lecithin. 


Hominy;  blood-flour;  bran-extract, 
(smaller  amount) 


Hominy;  blood-flour;  bone-meal 

Hominy;  blood-flour;  sodium  phosphate 
Check  lot 


Water 


78.21 

80.68 
78.80 

79.99 
79.27 
80.55 

78.80 


Protein 


15.78 

14.34 

16.38 

15.62 
15.49 
14.56 
16.83 


Fat 


3.07 

2.65 
2.89 

2.57 
2.94 
2.72 
1.99 


Ash 


1.17 

1.02 
1.15 

1.10 
1.11 
1.12 
1.20 


Phos- 
phorus 


.232 

.243 

.258 

.250 
.205 
.234 
.262- 


Propor- 
tion of 
phos- 
phorus 

to 
protein 


1.47 

1.69 
1.58 

1.60 
1.32 
1.61 
1.57 


Phos- 
phorus 
in  ash 


19.83 

23.82 
22.43 

22.73 
18.47 
20.90 
21.83 


EXPERIMENT  III 

1 

Con 

76.70 
80.32 
79.78 
77.18 
77.84 

17.07 
14.37 
15.79 
16.01 
17.00 

4.95 
2.11 
3.41 
3.92 
3.91 

1.14 
1.12 
1.11 
1.14 
1.07 

.313 
.249 
.249 

.318 
.294 

1.83 
1.73 
1.58 
1.99 
1.73 

27.46 

2 

Check  lot 

22.23 

3 
4 
5 

Hominy;  blood-flour;  bran-extract.... 
Hominy;  blood-flour;  bone-flour 

22.43 
27.90 
27.48 

DATA  CONCERNING  DEVELOPMENT  OF  BONES  (Forbes,  1909) 

EXPERIMENT  II 


Vol- 

Longer 

Shorter 

Lot 

Rations 

ume    of 
each 
humer- 
us 

each 
humer- 
us 

Ash 
per  c.  c. 

Breaking1 
strength 

Length 

trans- 
verse 
diam- 
eter 

trans- 
verse 
diam- 
-  eter 

C.  C. 

Grams 

Grams 

Lbs. 

Cm. 

Cm. 

Cm. 

1 

108.0 

111.8 
118.0 

117.6 

32.92 

33.33 
38.97 

36.39 

.3048 

.2981 
.3303 

.3094 

509 

575 
736 

676 

13.88 

13.68 
13.70 

13.74 

2.25 

2.23 
2.20 

2.34 

1.63 

? 

Hominy;  blood-flour;  bran-extract, 

1.67 

3 

1.66 

4 

Hominy;  blood-flour;  bran-extract, 

1.68 

5 

Hominy;  blood-flour;  bone-meal 

121.3 

46.22 

.3811 

791 

14.00 

2.28 

1.71 

6 

Hominy;  blood-flour;  sodium  phosphate 

112.5 

35.59 

.3164 

624 

13.90 

2.33 

1.69 

EXPERIMENT  III 


1 

74.6 
65.9 
100.1 
94.4 
93.7 

23.82 
19.93 
28.01 
28.97 
20.56 

.•319 

.3024 

.280 

.307 

.219 

502 
440 
641 
606 
426 

13.1 

11.64 

14.46 

13.46 

13.56 

1.83 
1.75 
1.98 
2.08 
1.95 

1.37 

9 

1.30 

3 
4 

R 

Hominy;  blood-flour;  bone-flour 

1.51 
1.46 
1.41 

PHOSPHORUS  METABOLISM 


375 


made  from  the  same.  Of  the  phosphorus  of  the  hay  53.4  percent 
was  used,  while  of  the  phosphorus  of  the  fresh  grass  91.0  percent 
was  used.  Fingerling,  therefore,  ascribes  the  difference  in  the 
value  of  the  phosphorus  in  cereals  and  mill  feeds,  on  the  one  hand, 
and  roughages,  on  the  other,  to  the  resistance  to  digestion  of  the 
crude  fiber  of  the  latter,  and  its  interference  with  the  digestion  of 
the  phosphorus  compounds. 

We  would  suggest  that  the  nature  and  amount  of  the  bases 
present  with  the  phosphorus  in  these  two  classes  of  foods  should 
receive  consideration  in  this  connection.  There  is  little  calcium  in 
the  cereals  and  mill  feeds,  and  much  more  in  roughage. 

Weiser  (1911, 1912)  has  studied  mineral  metabolism  with  swine 
on  cereal  diets,  with  and  without  additional  calcium.  The  balance 
data  are  as  follows : 


AVERAGE  DAILY  NITROGEN  AND  MINERAL  BALANCES  WITH  SWINE 

ON  RATIONS  OF  CEREAL  FOODS  WITH  AND  WITHOUT 

ADDITIONAL  CALCIUM— Grams 


Duration 
of  period 
in  days 

Rations 

Average 

body 
weight 

Kg. 

P 

Food 

Urine 

Feces 

Balance 

Ca 

Food 

Urine 

Feces 

Balance 

Mg 

Food 

Urine 

Feces 

Balance 

N 

Food 

Urine 

Feces 

Balance 

21 

55.88 

3.2073 

1.0241 

2.4841 

-0.3009 

0.2507 

0.1428 

1.3936 

-1.2857 

1.5868 

0.1951 

0.1206 

+1.2711 

21.39 
15.38 
3.19 

+2.82 

9 

53.37 

2.6731 

0.8134 

2.2570 

—0.3973 

0.1996 

0.1384 

1.1298 

—1.0686 

1.3201 

0.1337 

0.1276 

+1.0588 

17.82 

12.59 

2.67 

+2.66 

8 

44.12 

2.1467 

0.6663 

1.1986 

+0.2818 

0.1559 

0:0308 

0.3206 

-0.1955 

1.0593 

0.1623 

0.3878 

+0.5092 

14.32 

9.04 

2.13 

+3.15 

900  firm,  barley;  160  gm.  starch 

52.55 

2.5251 

0.7950 

1.2572 

+0.4729 

0.5621 

0.0489 

0.4934 

+0.0198 

1.1002 

0.2718 

0.7058 

+0.1226 

17.899 

8.810 

3.670 

+5.419 

10 

1050  gm.  corn;  5  gm.  calcium  car- 

47.02 

2.8167 

0.2714 

1.6960 

+0.8493 

2.1950 

0.0766 

1.2602 

+0.8582 

1.2528 

0.2075 

0.9440 

+0.1012 

18.78 

12.08 

2.36 

+    4.34 

7 

)00  gm.  barley;  3  gm.  calcium  car- 

49.93 

2.5251 

0.2862 

1.2238 

+1.0151 

2.1961 

0.0778 

0.6699 

+1.4484 

1.1009 

0.1962 

0.5968 

+0.3079 

17.760 

8.760 

3.310 

+5.690 

376  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Author's  Summary.  "1.  By  exclusive  feeding  on  corn,  growing 
swine  experience  a  calcium  and  phosphorus  deficit  even  when  there 
is  storage  of  flesh  and  fat.  At  the  same  time  there  is  storage  of 
magnesium. 

"2.  By  the  addition  of  calcium  carbonate,  not  only  was  the 
calcium  and  phosphorus  deficit  relieved,  but  there  was  even  a  good 
retention  of  calcium  and  phosphorus.  At  the  same  time  the  stor- 
age of  magnesium  was  decidedly  lowered.  The  amount  of  calcium 
carbonate  by  which  one  may  reckon  on  a  good  calcium  retention  is 
about  10-11  gm.  per  100  kg.  body  weight. 

"3.  From  what  has  been  said  it  is  evident  that  the  bone 
requirement  of  young  swine  is  not  sufficiently  met  by  a  feed  of  only 
corn  and  barley,  but  this  may  easily  be  helped  by  adding  calcium 
carbonate." 

Hagemann  (1912)  obtained  results  in  metabolism  experiments 
with  two  wethers  and  a  steer  showing  that  calcium  phosphate  is 
more  effective  to  increase  nitrogen  and  calcium  retention  when 
finely  milled  with  the  feed  than  when  simply  mixed  with  the  same 
at  the  time  of  feeding,  a  result  in  harmony  with  that  of  Herxheimer 
(1897),  who  found  18  gm.  of  calcium  carbonate  baked 
in  bread  more  effective  to  increase  the  solvent  power  of  the  urine 
for  uric  acid  than  30  gm.  of  the  same  fed  as  a  powder. 
These  results  suggest  that  the  mechanical  relations  of  food  constitu- 
ents may  affect  their  time,  and  place  of  digestion,  and,  in  this  way, 
their  final  disposition.  See  also  Landsteiner  (1892) ;  effects  of  hay 
and  milk  on  blood-ash  of  rabbits. 

E.  B.  Forbes  and  associates  (1914)  have  shown,  in  their  experi- 
ments of  Series  IV,  that  corn  in  the  diet  of  swine  has  definite 
specific  effects  on  various  items  of  composition,  among  these, — the 
total  and  inorganic  phosphorus  of  the  blood,  the  lecithin  content  of 
the  liver,  and  the  composition  of  the  ash  of  the  bones. 

Forbes,  Beegle,  Fritz  and  Mensching  (1914)  conducted  an 
extensive  mineral  metabolism  experiment  with  swine,  phosphorus 
balances  being  included.  Five  barrow  pigs,  about  6  months  old, 
and  all  from  the  same  litter,  were  the  subjects  of  the  experiment. 

Confined  in  metabolism  crates  they  were  taken  through  eight 
10-day  collection  periods,  separated  by  7-day  intervals  on  the  next 
ration  to  follow,  the  change  of  food  being  made  abruptly  at  the  end 
of  the  collection  period.  The  five  animals  were  given  the  same  kind 
of  food,  the  results  therefore  being  based  on  five  repeats. 

The  foods  used  in  the  several  periods  were  as  follows : 

1.  Corn.  5.     Corn;  meat  meal. 

2.  Corn ;  soy  beans.  6.     Corn ;  skim  milk. 

3.  Corn;  linseed  oil  meal.  7.     Corn. 

4.  Corn ;  wheat  middlings.  8.     Rice  polish ;  wheat  bran 


PHOSPHORUS  METABOLISM  377 

They  were,  therefore,  the  common  practical  foods  for  swine  in 
this  country,  except  for  ration  No.  8,  composed  of  rice  polish  and 
wheat  bran,  these  feeds  being  selected  on  account  of  their  very  high 
content  of  magnesium  as  compared  with  calcium.  Corn  was  fed 
alone  in  the  first  and  seventh  periods  to  show  any  such  changes  as 
might  be  due  to  the  long-continued  routine  or  to  increasing  age. 

The  observations  covered  the  usual  proximate  analyses  of  food- 
stuffs and  feces,  daily  nitrogen,  creatinin  and  ammonia  estimations 
on  the  urine,  also  determinations  of  sodium,  potassium,  calcium, 
magnesium,  sulphur,  phosphorus  and  chlorine  on  foods,  urines  and 
feces;  and  further,  a  slaughter  test  on  the  five  animals  after  the 
termination  of  the  experiment. 

The  condensed  mineral  balance  data  are  in  the  table  on  p.  378. 
These  figures  represent  averages  of  results  from  five  individuals. 
The  uniformity  of  the  results  with  the  several  individuals  was 
sufficiently  marked  to  warrant  averaging. 

The  phosphorus  balances  in  these  experiments  were  all  positive, 
except  for  one  individual  on  the  ration  of  corn  alone.  In  no  case, 
however,  was  there  any  considerable  retention  of  phosphorus  on 
corn  alone.  Except  in  one  case  the  phosphorus  retention  in  the  sev- 
eral periods  was  in  the  same  order  as  the  intake.  This  exception 
was  the  ration  containing  the  wheat  middlings.  The  peculiarity 
of  the  phosphorus  of  this  ration  was  a  large  proportion  of  tritico- 
nucleic  acid,  ands  the  phosphorus  of  this  ration  was  much  less 
efficiently  retained  than  the  phosphorus  of  the  rations  containing 
meat  meal  and  milk.  With  a  much  smaller  intake  of  phosphorus  in 
meat  meal  and  in  milk  the  retention  was  much  greater.  Two  circum- 
stances unfavorable  to  phosphorus  retention  in  the  wheat  middlings 
ration  were  the  presence  of  much  less  calcium  and  much  more  mag- 
nesium than  in  the  meat  meal  and  milk  rations.  The  results  were 
increase  in  both  urine  and  feces  phosphorus. 

The  magnesium  of  the  food  is  shown  to  be  a  prominent  factor 
in  the  partition  of  the  phosphorus  between  urine  and  feces,  an 
increased  proportion  of  magnesium  to  phosphorus  in  the  food 
increasing  the  proportion  of  feces  to  urine  phosphorus.  There  was 
no  such  prominent  effect  of  magnesium  to  restrict  phosphorus  reten- 
tion. 

During  the  feeding  of  skim  milk  there  were  lower  proportions 
of  potassium,  magnesium,  sulphur,  chlorine  and  phosphorus  in  the 
feces  than  during  any  other  period ;  and  this  period  was  also  charac- 
terized by  the  maximum  percentage  retention  of  the  calcium,  magne- 
sium, sulphur  and  phosphorus  intake. 


S78 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


s 

o 

fa 

fa 

Q 

fa 

cc 

O 

fa 

o 

JO 

U 

I-H 

CT2 

«5 

rH 

& 

7) 

O 

t— i 

H 

s 

- 

S§ 

ft 

a 

0/ 

I 

■a. 

K° 

- 

°Z 

«d 

fa 

So 

s 

01 

> 

fa 

i^'d 

s- 

«jgfa 

0 

r  o 
fau 

<£ 

0. 

r.f 

o 

if.  -, 

S 

fa 

S- 

3 

> 

i— i 

<l 

fa 

0) 

o 

,o 

fa 

•- 

o 

<U 

fa 

H 

fa 

s 

fa 

«! 

fa 

fa 

fc 

<MC5t^i>-  cmcooc-h  cococoos  o  — <~-<i>-  mt—  <mco  cor-oso  ooocc-* 
Wf-in-i  io^-r—;o  xci^o  car—THO  cMCor^CD  CDOor-ai  cciot"--* 
c^  —  c:  t~-      coco  co  co      (nxicsi      tji  cd  i>-  o      cx>  cm  oc  i>-      nho:o      tp  cg  m  o 


r^COCD'— l        •-t(Nt>i> 


t— (Ot*-CM 


t-HOTf>H 


+ 


+ 


■gcSc 

\8-cB5 

feP&« 


^i3^*^Oi      — <  cm  ro  t—      lO-^-rtiGO      -rt<ir>T#iT*      co  r--  oi  co      i>-  ct>  cm  *n>      co  co  :o  cm 
c:  :d  cm  :d      cc  m  cm  o      co  m  cm  io      t*  cm  o  t— i      co  os  m  o      ccjdoco      ocooa  oa 


fl  CMlOCO        lO-— I  SSlO        COI>-lO 


r-^'sicooo     iocdoco     c*-ooo505 


IOHMO        CDt-i^O 


or8-cai3 

Effort 


«r8-cSiS 


•rt  "  oi  ~ 

fepk£ 


»i « oi  y 

rt  o  c  s  E 
or8-c8  5 

kpfepg. 


rt  o.S  8  « 
^fcpk  rt 


^   I)   01   J! 

W  811.2 

f^pfcrt 


a  «  "  ^.^f 


Ah 


^Ot^CD        ffiONh-        CDOCjOOCC) 


GOO^r-D        CO  CO  -^  Oi        WOj^O) 


i -  -^  ? i  ^      ro  cc  ■;f  :d      io  *— i  co  oi      oincc'*      ^iihoi      oo^^      irci  o  m  co 

t-OOOJ        t^  CO  O  O        1<OH!N        CO  t—  O  00        HOOW        OIOO"*        CM  t^  O  T*. 


+ 


IOIOOO        020000        OCCOO.-1        lOTKOO        1DCOOH 

+         +         +         +        + 


r-^, — i  r-vj-r^        Oi  t-inCO        r*i  00  CO  00        »— <  CO  **  rtf        t—  CO  O  CO        tM  t— I  **t«  CO        CO  <M  CM  CO 

:,^:]-ti       i.oop'r       rc;7.a       re  :d  cm  -*      -ficocoo      t^ooco      cdo^p: 


CMIQCDO       r-CDCCCO       cccooco       coho-* 


•*^lO       lOTtl^t-       coocoo 


+ 


+ 


COt— It— li— I         COHOH         NHOO         PJHOH 

+         +         +         + 


i— ICMCMCO        COOCOCO        -*-t<COCt>tO        »— It- 

t*  cm  o  "*      cd  —  -r  CO      iocm -rn t--      m  ^* 


^        QOO^^        COCOf-CD        COCDOt-h 

cocmojcm     cct^oarH     r*  co  oo  ^i 


— ^  5a  c.  cd      co io 03 o      -rinoccD      t— icoosi—t      co-^cch      occ^r— i— i      oj-^iot^- 


NOHO      mowo 

+  I 


iOr-lO        OJOOiQ 


XOIOM        OOJh-^l        COC^OCCO        CO 


oocDinr-     conon     cocd»-400 


faC~,OT*  XOM-t  CDOOOiOD  CO »— )  -^  CO  O3C0I>--n        OICOO^CM        t^rtCOO) 

t>-od^CV)  COT-HCiCO  QrHCDO  CMCOt-CM  CD-HCOO        CMi-^lOTtl        I>- «— I  — I  -rji 

dddd  T-HCDcJd  ddoo  ioot-ico 

I  +  +  + 


lC<3        OOOO 


rtmfw 


inawt-     oa?no 


j^i^fo     iocoi^»n     oo-~m»— i     ccr~meo     Oifocot^     iofo-^co 


COC^liOlO        T-H-Ht-*<-H        CO^^^-^I 


WO        OH  —  CCOO        ODNOO        t—  03COT«        CXjTft^CO 


30>CM        lOt— (t^-C~-        O^CDlOtM        »— ICSIOCO 


1C<l»-iT— <        COIOHO 


IMCOOO        TJtr-l— l-^ 


+ 


r-  cr.  en  co      cc  io 

X)  QO  CO  CM       CD  t- 

—  r^  uo  oo     t— 1 1*- 


T-^OOCOiO       COr*-OJ00       t-HIOC»CO       Od^MC^OQ       i— KMIOCO 

^t^oc:      I-—  -r  mcj      r— ^t^-t^-      oa  mem      c^jecr-oo      cc  co «— i  cs 

TPOmX        (XHlCH        t— IOCDTJ1        O5CC0Ct-1        TjiCDCNC^ 


H     T 


C-3  CO  -» i— I        CO  TJ<  I— I  rH 

rt     +         + 


fvjx* 11^-^H        rKr-(ION        t~05CM-t<        COinCOOO        CNCMOCM  CM  O  CO  t— (r-.COCM 

t^^H^-5!     ^cjo-^cm     or^coo     incsc^t--     occococo        accoi—       oiocccsj 


.IO        rlCO      -IO        OI 


QOCO      -CM 


E01  *j  ^       p 
Oorti>       o-r  rt^,       °>rt|> 

OKCCr^         OrJWt?         OtSCC^ 


BJa- 


:  c 

*  rt 

01  b 

—  *° 

P-rt 

85^15 


CO  t^ 


CO  CO 


CO  03 


PHOSPHORUS  METABOLISM  379 

Corn  is  shown  to  be  more  deficient  in  calcium  than  in  any  other 
nutrient;  its  magnesium  content  is  also  low,  and  its  phosphorus 
content  allows  of  but  slight  retention.  At  the  same  time  the  nitro- 
gen retention  is  quite  considerable.  In  spite  of  the  slight  retention 
of  phosphorus  its  amount  was  considered  as  insufficient,  since  there 
was  present  hardly  more  than  the  amount  required  for  maintenance, 
at  a  time  of  life  which  would  naturally  be  characterized  by  exten- 
sive storage  of  phosphorus. 

The  results,  in  general,  show  that  the  mineral  requirements  of 
swine  are  apt  not  to  be  satisfied  during  cereal  feeding.  A  dry-lot 
fattening  process  probably  involves,  as  a  rule,  considerable  draft 
upon  mineral  stores  previously  accumulated  during  periods  of  access 
to  green  feeds. 

In  a  series  of  experiments  by  Forbes  and  associates  (1914),  with 
swine,  three  lots  of  pigs  (5  in  a  lot),  receiving  corn  in  the  ration, 
differed  as  a  group  from  five  other  lots  which  received  no  corn  in  the 
ration,  in  several  respects,  as  follows :  Each  of  the  three  lots  of  pigs 
receiving  corn  had  more  potassium  and  magnesium,  and  more  total 
and  inorganic  phosphorus  in  the  blood,  more  magnesium  and  phos- 
phorus, and  less  calcium  in  the  bone  ash,  and  more  lecithin  in  the 
liver  than  any  one  of  the  five  lots  which  had  not  received  corn. 

For  a  summary  on  common  foods  in  relation  to  phosphorus 
metabolism  see  p.  395.  See  also  the  following  sections  on  the 
phosphorus  content  of  milk,  and  the  phosphates  of  the  bones,  as 
affected  by  foods. 

THE  PHOSPHORUS  CONTENT  OF  MILK  AS  AFFECTED  BY  FOODS 

The  idea  of  modifying  the  composition,  and  thus  the  nutritive 
value  of  milk,  through  the  character  of  the  food,  is  ancient,  and  has 
been  the  inspiration  of  many  investigations.  Some  misconceptions 
among  the  conclusions  reached  have  been  due  to  the  relatively 
considerable  variations  in  the  composition  of  the  milk  during  the 
course  of  the  period  of  lactation. 

Among  the  earlier  observations  were  those  of  Bocker  (1849) 
who  stated  that  administration  of  calcium  phosphate  to  wet  nurses 
whose  milk  was  poor  in  lime  served  to  enrich  the  milk  in  this  constit- 
uent; and  Ssubotin  (1866),  who  studied  the  composition  of  dog's 
milk  as  affected  by  various  rations,  and  ascribed  an  increased  casein 
content  to  fat  in  the  diet;  also  Decaisne  (1871)  who  made  observa- 
tions on  43  lactating  women  during  the  siege  of  Paris.  Hunger 
prevailed;  the  flow  of  milk  was  often  insufficient  to  nourish  the 
infants,  and  12  died  of  starvation.  The  milk  was  low  in  casein,  fat, 
sugar  and  salts,  but  high  in  albumin. 


380 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


Weiske  (1871b)  studied  the  effects,  on  the  composition  of  cow's 
milk,  of  adding  20  grams  daily  of  acid  calcium  phosphate  to  a  ration 
which  the  cows  had  been  consuming  regularly  during  a  protracted 
period.  There  were  no  differences  caused  by  the  use  of  calcium 
phosphate,  nor  did  Yvon  (1879)  succeed  in  increasing  the  phosphate 
of  cow's  milk  in  this  way;  but  Hess  and  Schaffer  (1891)  claim  that 
they  did  produce  increased  phosphorus  in  the  milk  ash.  The  aver- 
age percent  of  P205  in  the  ash  of  the  milk  of  three  cows  before  feed- 
ing the  phosphate  was  25.80 ;  after  about  a  month  this  figure  was 
increased  to  27.52,  and  after  about  two  months  to  29.54 ;  but  it  was 
not  shown  by  the  use  of  controls  that  the  advance  in  the  period  of 
lactation  did  not  enter  into  the  production  of  this  result.  Sagnier 
(1891)  reports  that  Charles  Gravier,  by  special  feeding  (the  nature 
of  the  feeding  is  not  revealed),  was  able  to  furnish  for  the  hospitals 
milk  containing  from  2.30  to  2.50  gms.  phosphoric  acid  per  liter. 
Duclaux  (1893)  concludes  that  milk  which  is  sold  as  rich  in  phos- 
phate, because  of  phosphate  feeding,  is  found  not  to  be  so  on  anal- 
ysis. 

Hills's  experiments  (1887,  1894)  with  ground-bone  feeding 
showed,  first,  that  the  added  phosphorus  did  not  all  appear  in  the 
feces ;  then  that  it  increased  somewhat  the  phosphoric  acid  content 
of  the  milk  of  the  cows ;  in  one  case  from  0.2142  to  0.2263  percent, 
and  in  another  from  0.1809  to  0.1909  percent. 

J.  Neumann  (1893a)  reports  an  experiment  in  which  the  addi- 
tion of  a  calcium  phosphate  preparation  to  a  feed  already  supplying 
sufficient  quantities  of  the  mineral  constituents  for  milk  production 
seemed  to  cause  a  slight  rise  in  the  lime  and  phosphoric  acid  content 


LIME  AND  PHOSPHORIC  ACID  OF  MILK  AS  AFFECTED  BY  CALCIUM 
PHOSPHATE  ADDED  TO  THE  FEED  (J.  Neumann,  1893a) 


Milk 

In  1000  gm.  milk 

Percent  of  ash 

In  gms.  per  cow 

Date 

Kg. 

Ash 
Percent 

Lime 

Phos- 
phoric 
acid 

Lime 

Phos- 
phoric 
acid 

Lime 

Phos- 
phoric 
acid 

Aug-.  22 
"     23 
"     24 

27.16 
27.40 
28.20 

0.77 
0.77 
0.77 

1.4950 
1.4804 
1.4618 

1.9271 
1.9594 
1.9930 

19.42 
19.23 

18.98 

25.03 
25.45 
25.88 

13.53 
13.51 
13.74 

17.44 
17.88 
18.73 

Without  added  phos- 
phate: about  80  gm.  lime, 
and  110  gm.  phosphoric 
acid  per  day  per  cow 

"    27 

"     31 

Sept.  3 
r.     7 

"     13 
"     20 
"     28 

24.85 
25.79 
25.67 
25.00 
25.50 
24.30 
24.12 

0.75 
0.75 
0.78 
0.78 

0.'77 
0.77 

1.4355 
1.4371 
1.5566 
1.4995 
1.5252 
1.5916 
1.5509 

1.8865 
1.9307 
1.9864 
1.9868 
1.9685 
2.0262 
2.1323 

19.14 
19.16 
19.96 
19.22 
(19.81) 
20.67 
20.14 

25.15 
25.74 
25.47 
25.47 
(25.56) 
26.31 
27.69 

11.88 
12.36 
13.32 
12.49 
12.96 
12.90 
12.47 

15.62 
16.61 
17.00 
16.55 
16.73 
16.41 
17.14 

40  grm.  lime  and  33  g-m. 
phosphoric  acid  per  day 
per  cow  added  as  calcium 
phosphate 

PHOSPHORUS  METABOLISM 


381 


of  the  milk,  but  only  slight,  and  only  after  three  or  four  weeks' 
duration  of  the  phosphate  feeding.  The  first  effect  of  the  phos- 
phate was  suddenly  to  reduce  the  milk  flow.  The  milk  of  three 
cows  was  mixed  for  the  analyses  quoted  on  p.  380.  The  first  addition 
of  phosphate  was  made  on  the  evening  of  August  24. 

Sanson  (1894)  reports  the  following  results,  which  were  con- 
sidered to  show  increasing  amounts  of  phosphorus  in  milk,  associ- 
ated with  increasing  amounts  of  Na2HP04  added  to  the  feed,  up  to 
the  dose  of  22  gms.,  no  further  increase  occurring  with  greater 
amounts  of  the  phosphate.  The  increased  phosphorus  of  the  milk 
was  all  in  the  serum,  the  casein  not  showing  any  increase. 

PHOSPHORUS  OF  MILK  INCREASED  BY  PHOSPHATE  FEEDING 

Sanson  (1894) 


Day 

Dose  of  sodium  phosphate 

Density  of  milk 

P2O5  of  milk 

Increase  of  P2O5 

Grams 

Grams  per  1000 

Grams  per  1000 

1 

0 

1.032 

Mi          1-438 

0 

2 

10 

1.031 

1.984 

0.546 

3 

.12     ' 

1.033 

2.046 

0.608 

4 

14 

1.031 

1.946 

0.508 

5 

16 

1.029 

2.046 

0.608 

6 

18 

1.032 

2.084 

0.646 

7 

20 

1.032 

2.145 

0.707 

8 

22 

1.036 

2.170 

0.732 

9 

24 

1.034 

2.070 

0.632 

10 

26 

1.037 

2.108 

0.670 

11 

28 

1.034 

2.083 

0.645 

12 

30     . 

1.033 

2.170 

0.732 

In  the  experiments  of  Jordan,  Hart  and  Patten  (1906),  with 
two  cows,  the  removal  of  the  phytin  from  the  bran  fed  was  with- 
out significant  change  on  the  phosphorus  content  of  the  milk. 

EFFECTS  OF  THE  INGESTION  OF  PHOSPHORUS  COMPOUNDS  ON  THE 

COMPOSITION  OF  MILK— MEAN  DAILY  OUTPUT  IN  MILK 

Jordan,  Hart  and  Patten  (1906) 


Composition  of  milk,  percent 

Partition  of  phosphorus  in  milk 

Dates 

Pro- 
teids 

Case- 
in 

Fat 

Sugar 

Solids 

Total 
Gms. 

Nucleo- 
phos- 

phorus 
Gms. 

Soluble 

org-anic 

Gms. 

Inor- 
ganic 
Gms. 

Ration 

Mar.  12-Mar.  18 
Mar.  30-Apr.  5 

Apr.  24-May  1 
May  29- June  4 

3.07 
3.05 

3.22 
3.27 

2.53 
2.45 

2.66 
2.75 

3.28 
3.09 

3.73 
3.29 

5.56 
5.46 

5.47 
5.50 

11.91 
11.59 

12.41 

12.07 

14.7 
15.8 

3.5 
3.3 

1.05 
1.17 

10.2 
11.2 

Oat  straw,  bran, 
rice  meal,  wheat 
gluten. 

Oat  straw,  washed 
bran,  rice  meal, 
larger  amount  of 
gluten. 

Lipschitz  (1906)  notes  that  the  feeding  of  bone  meal  reduces 
the  milk  flow,  though  increasing  the  total  ash  content  of  the  milk, 


382 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


Hart,  McCollum  and  Humphrey's  observations  (1909)  on  one 
cow,  extending  over  three  and  a  half  months,  are  summed  up,  so  far 
as  the  ash  of  the  milk  is  concerned,  by  saying :  "Variations,  within 
wide  limits,  in  the  form  and  quantity  of  supply  of  potassium,  mag- 
nesium, or  phosphorus  did  not  influence  the  percentage  content  of 
these  elements  in  the  milk."  It  is  to  be  noticed  that  the  amounts 
yielded  fell  off  markedly  during  the  experiment. 

Von  Wendt's  (1909)  conclusion  from  his  work  was  that  neither 
the  fodder  nor  the  added  salts  exerted  much  influence  on  the  compo- 
sition of  the  milk,  the  phosphorus,  casein  and  calcium  being  among 
the  items  least  affected  of  all. 

CHANGES  OF  AMOUNT  AND  COMPOSITION  OF  MILK  ACCOMPANYING 
THE  ADMINISTRATION  OF  CERTAIN  SALTS  (Von  Wendt,  1909) 


Gain  or  loss  of  P2O5 

Gain  or  loss  of  CaO 

Dates 

Supplements 

Sain  or  loss 
in  milk 

Cow  or 

| 

group 

Grams 

Percent 

Grams 

Percent 

Grams 

Oct.  14-16 

30  gm.  potassium  oxalate 

—2000 

—0.002 

-2.3 

-0.02 

-4.8 

Oct.  20 

60  gm.  potassium  oxalate 

+900 

0 

+0.9 

-0.03 

—2.8 

Nov.  2-3 

60  gm,  potassium  oxalate 

—770 

+0.006 

+0.011 

+0.006 

+0.3 

Nov.  19-21 

75  kg.  beet  leaves 

—2150 

+0.001 

—2.1 

0 

-2.8 

Cow  149 

Dec.  16-18 

30  gm.  Ca  HP04 

+480 

+0.002 

+0.7 

+0.042 

+5.1 

Cow  151 

May  26-28 

75  gm.  Ca  HPO4 

-300 

-0.007 

-1.1 

+0.026 

+2.9 

Cow  150 

May  26-28 

75  gm.  Ca  HPO4 

—300 

-0.003 

-0.7 

+0.024 

+3. 

Cow  150 

Dec.  16-18 

30  gm.  Ca  HPO4 

-600 

+0.001 

-0.7 

+0.013 

0 

Cow  149 

Dec.  25-27 

90  gm.  lime 

+330 

0 

+0.35 

-0.007 

-0.25 

Cow  150 

Dec.  25-27 

90  gm.  lime 

—400 

-0.001 

-0.23? 

+0.021 

+0.7 

Cow  150 

June  29- July  4 

15  gm.  lecithin 

—1300 

+0.004 

—0.8 

+0.020 

+1.1 

Gr.     I 

Apr.  22-24 

30  gm.  Ca  HPO4 

+100 

+0.002 

+0.45 

+0.007 

+1.4 

Gr.   II 

Apr.  22-24 

30  gm.  Ca  HPO4 

"-200 

+0.001 

-0.1 

+0.006 

+0.27 

Gr.  Ill 

Dec.  7-9 

90  gm.  CaHP04 

+700 

+0.001 

+0.66 

0.0 

+0.63 

Gr.    II 

Dec.  25-27 

60  gm.  Ca  HPO4 

-220 

—0.004 

—0.2 

-0.007 

-1.0 

Gr.  Ill 

Dec  28-30 

60  gm.  Ca  HPO4 

+400 

+0.001 

—0.06 

+0.006 

-1.0 

Gr.     I 

Apr.  16-18 

30  gm.  lime 

+1400 

-0.001 

+1 

0 

+2 

Gr.    II 

Apr.  16-18 

30  gm.  lime 

—500 

-0.001 

-0.57 

-0.006 

-1.18 

Gr.     I 

Dec.  28-30 

90  gm.  lime 

+400 

+0.004 

+0.9 

+0.014 

+2.3 

Gr.    II 

Dec.  16-18 

90  gm.  lime 

+40    ' 

+0.001 

+0.2 

+0.042 

+5 

Gr.  Ill 

Dec.  7-9 

90  gm.  lime 

-110 

+0.003 

+0.15 

+0.007 

+0.5 

Gr.   II 

Oct.  19-21 

Beet  leaves+Ca  HPO4 

—460 

-0.001 

-0.6 

0 

-0.6 

Gr.  Ill 

Oct.  19-21 

Beet  leaves+Ca  HPO4 

—440 

0 

-0.4 

0 

-0.5 

Gr.  Ill 

Nov.  29-Dec.  6 

Beet  leaves+Ca  HPO4 

+900 

+0.009 

+2 

+0.013 

+2.2 

Golding  and  Paine  (1910)  planned  an  experiment  to  test  the 
possible  effects  on  milk  of  pasturing  cows  on  land  manured  with 
phosphates  and  potash,  as  compared  with  pasturing  on  unmanured 
land  on  grass  of  very  poor  quality.  The  ash,  phosphoric  acid, 
potash  and  lime  of  the  milk  (averaged)  are  the  same  for  the  two 
plots  of  land.  The  milk  yield  was  considerably  higher  from  the 
cows  on  the  manured  half,  and  its  fat  content  higher  from  the  other 
half. 

According  to  the  data  furnished  by  Monvoisin  (1910),  both, 
inflammation  of  the  udder  and  tuberculosis  affect  the  composition  of 
the  milk,  lowering  the  percent  of  phosphorus  in  the  ash.  His  data 
also  show  a  distinct  lowering  of  the  casein  content  of  milk  in  tuber- 
culosis, both  in  absolute  amount  and  in  relation  to  total  nitrogenous 
matter. 


PHOSPHORUS  METABOLISM 


383 


A.  R.  Rose  and  J.  T.  Cusick  (1911)  report  inconclusive  studies 
of  the  influence  of  phosphorus  of  the  food  on  the  yield  and  composi- 
tion of  goat  milk. 

Fingerling  (1911b)  studied  the  effects  of  diets  rich  and  poor  in 
calcium  and  phosphorus,  and  of  dicalcium  phosphate  and  calcium 
carbonate,  on  the  yield  and  composition  of  the  milk  of  goats;  and 
also  the  effects  of  the  same  on  the  calcium  and  phosphorus  balances. 
The  balances  responded  quickly  and  consistently  to  variations  in  the 
intake  of  calcium  and  phosphorus.  The  effects  of  these  conditions 
on  the  composition  of  the  milk  were  slight;  the  effects  on  the 
amount  of  milk  produced  were  more  prominent. 

INFLUENCE  OF  A  LIME-  AND  PHOSPHORIC  ACID-POOR  DIET  ON  THE 
MILK  SECRETION  OF  GOATS  (Fingerling,  1911b) 


Amount  of  milk 

Ash 

Percent 

of  ash 

No.  of 

Goat 

Feed 

days 

Grams 

Grams 

CaO 

P2O5 

A 

Lime-  and  phosphoric  acid-rich 

1958 

13.31 

18.09 

25.89 

14 

Lime-  and  phosphoric  acid-poor 

1886 

13.30 

18.04 

25.80 

14 

Lime  and  phosphoric  acid 

1547 

10.71 

19.79 

29.85 

13 

Lime   and  phosphoric  acid 

1388 

8.90 

21.44 

30.41 

13 

Same  with  dicalcium  phosphate 

1493 

10.35 

19.97 

28.35 

14 

Same  with  dicalcium  phosphate  and  CaC03 

1687 

11.72 

18.68 

26.41 

17 

" 

Like  first  period 

1958 

13.31 

18.09 

25.89 

31 

Lime-  and  phosphoric  acid-rich 

1011 

8.59 

23.06 

29.89 

13 

Lime-  and  phosphoric  acid-poor 

999 

8.66 

23.23 

29.51 

14 

Lime-  and  phosphoric  acid-poor 

839 

7.08 

24.22 

33.43 

14 

Lime-  and  phosphoric  acid-rich 

787 

6.55 

26.20 

35.35 

12 

Lime-  and  phosphoric  acid-poor 

1061 

8.73 

22.63 

30.39 

12 

Lime-  and  phosphoric  acid-rich 

1011 

8.59 

23.06 

29.89 

9 

From  later  experiments  it  was  concluded  (Fingerling,  1912b) 
that  neither  the  activity  of  the  mammary  glands  nor  the  composi- 
tion of  the  milk  was  altered  by  feeding  the  organic  phosphorus 
compounds. 

EFFECTS  ON  GOAT'S  MILK  OF  ADDING  DIFFERENT  PHOSPHORUS 
COMPOUNDS  TO  A  BASAL  RATION  VERY  POOR  IN  PHOSPHORUS 

Fingerling    (1912b) — Grams 


Ration 

Milk 

CaO 

P2O5 

Date 

Total 

Gain  or  loss 

Total 

Gain  or  loss 

Total 

Gain  or 
loss 

A 
B 

A 
B 

A 
B 

A 
B 

A 

A 

A 
B 

Basal  ration 
Basal  ration 

Lecithin  added 
Lecithin  added 

Phytin  added 
Phytin  added 

Casein  added 
Casein  added 

Nuclein  added 

Sodium  nucleate  added 

Di-sodium  phosphate  added 
Di-sodium  phosphate  added 

1587 
1396 

1515 

1563 

1397 
1593 

1525 
1606 

1491 

1430 

1583 
1824 

—72 
+167 

—190 
+197 

-62 
+210 

-96 

-157 

-4 
+428 

1.808 
3.060 

1.838 
3.072 

1.813 
3.136 

1.784 
3.083 

1.828 

1.804 

1.883 
3.179 

+0.030 
+0.012 

+0.005 
+0.076 

-0.024 
+0.023 

+0.020 

-0.004 

+0.075 
+0.119 

2.594 
4.164 

2.626 
4.247 

2.555 
4.351 

2.531 
4.255 

2.595 

2.561 

2.656 
4.401 

+0.032 

+0.083 

-0.039 
+0.187 

-0.063 
+0.091 

+0.001 

-0.033 

+0.062 
+0.237 

384  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Gaube  (1895)  has  also  studied  this  question,  and  Jensen  (1904, 
1905a,  1905b,  1905-6)  has  made  extended  investigations  in  this  field, 
especially  with  rations  of  varying  proportions  of  hay  and  beets  to 
which  were  added  phosphates  of  sodium,  calcium  and  magnesium, 
etc.  Lanzoni  (1918)  noted  slight  modifications  of  composition  of 
milk  through  the  agency  of  drugs.      See  also  Vivier  (1911). 

In  all  cases  the  variations  in  phosphorus  content  of  cow's  milk 
which  are  ascribable  to  the  character  of  the  food  are  within  the 
range  of  normal  variation.  At  most  these  differences  are  slight, 
or  doubtful,  and  in  no  case  of  practical  significance  from  any  point  of 
view. 

As  to  the  effects  of  starvation — the  evidence  is  insufficient  to 
warrant  conclusions. 

EFFECTS  OF  DIET  ON  THE  PHOSPHATES  OF  THE  BONES 

As  the  principal  store  of  phosphates  in  the  body  the  skeleton 
is  of  interest  in  connection  with  the  various  circumstances  which 
affect  phosphorus  metabolism.  In  judging  of  the  results  of  experi- 
mental feeding,  however,  we  must  bear  in  mind  the  fact  that  the 
amount  of  phosphorus  in  the  bones  is  so  very  much  greater  than 
that  in  the  remainder  of  the  body  that  we  may  be  unable  to  detect 
changes  of  composition  in  the  skeleton  under  the  influence  of 
circumstances  which  produce  marked  effects  on  phosphorus  metabo- 
lism as  revealed  by  balance  experiments.  The  skeleton  is  so  vari- 
able in  its  development ;  its  mineral  constituents  are  so  much  affect- 
ed by  the  food,  that  special  attention  must  be  given  to  the  breeding 
and  rearing  of  experimental  subjects  for  comparative  studies  on  the 
bones.  In  our  own  experiments  it  has  been  found  possible  greatly 
to  decrease  the  central  cavity  in  the  shaft  of  the  long  bones  of  pigs 
by  feeding  rations  which  are  rich  in  bone-forming  constituents. 
There  seems  to  be  no  definite  upper  limit  of  phosphate  deposit  in  the 
bones,  as  there  is  in  the  soft  parts. 

Chossat  (1842)  found  that  pigeons  fed  on  wheat  alone  died  in 
8-10  months  from  malnutrition.  The  salts  were  gradually  with- 
drawn from  the  bones,  which  became  weakened,  and  the  breast  bone 
imperforate.  The  addition  to  the  diet,  of  calcium  carbonate, 
appeared  to  prevent  these  symptoms,  and  to  maintain  the  birds  in 
health.  Apparently  the  lack  of  calcium  limited  the  use  of  phos- 
phorus. In  a  later  paper  Chossat  (1843)  makes  a  full  report  of  his 
experiments  with  pigeons,  chickens,  rabbits,  frogs,  lizards,  eels, 
tortoises,  and  other  animals.  The  conclusions  were  in  harmony 
with  the  above.  Where  pigeons  lost  40.4  percent  of  the  live  weight 
in  starving  to  death,  only  3.4  percent  of  this  loss  seemed  to  come 
from  the  bony  system. 


PHOSPHOKUS  METABOLISM  385 

Among  other  early  studies  are  those  of  Boussingault  (1846a, 
1846b),  who  investigated  the  effects  of  foods  on  the  growth  of  the 
skeleton  of  pigs,  and  C.  Falck  (1850),  who  studied  the  effects  of 
feeding  lime-rich  and  lime-poor  foods  on  the  development  of  domes- 
tic fowls. 

Dusart  (1869)  determined  that  a  pigeon  lost  4.9  gm.  of  calcium 
phosphate  from  the  skeleton,  and  58  gm.  of  live  weight  during  83 
days'  feeding  on  wheat.  The  addition  of  calcium  carbonate  to  this 
diet  reduced  to  zero  the  alkali  phosphates  of  the  excreta,  which  had 
been  half  of  the  total,  and  caused  a  return  to  the  normal  live  weight. 

Dusart  (1870)  also  published  clinical  reports  of  19  hospital 
cases  of  fracture  or  disease  of  the  bones,  under  treatment  with  cal- 
cium lactophosphate,  which  he  recommends. 

Weiske  (1871a),  in  experiments  with  milch-goats,  found  that 
where  the  food  was  especially  poor  in  calcium  and  phosphorus,  or  in 
calcium  alone,  death  might  be  caused  by  these  deficiencies  before 
noticeable  change  was  produced  in  the  composition  of  the  bones. 
Probably  greater  refinement  of  experimental  procedure  would  have 
demonstrated  some  change  in  the  bones. 

Weiske  found  (1872a)  that  calcium,  magnesium  and  strontium 
phosphates,  when  fed  to  rabbits  with  a  ration  of  hay  and  turnips, 
produced  no  important  change  in  the  composition  of  the  ash  of  the 
bones. 

Weiske  and  Wildt  (1873)  fed  one  lamb  on  a  normal  ration,  and 
two  more  on  a  basal  ration  of  straw,  starch,  sugar  and  casein,  which 
was  low  in  calcium  and  phosphorus.  One  of  these  also  received 
calcium  carbonate  and  the  other  disodium  phosphate.  The  experi- 
ment lasted  25  days.  The  basal  ration  seems  to  have  contained 
enough  calcium  and  phosphorus  so  that  the  experimental  procedure 
produced  in  25  days  no  consistent  change  in  the  composition  of  the 
bones. 

Hofmeister  (1873)  experimented  with  8  lambs,  4  in  each  of  two 
lots,  with  a  low-phosphorus  ration  of  meadow  hay  and  potatoes,  to 
which  he  added,  with  one  lot,  bone  phosphates  as  precipitated  from 
a  hydrochloric  acid  solution  by  sodium  carbonate.  The  experiment 
extended  from  June  7-Nov.  30. 

The  calcium  phosphate  caused  no  appreciable  increase  in  rate 
of  gain  in  weight.  Hofmeister  considered*  that  he  had  proven  the 
digestibility  of  precipitated  phosphates  by  lambs,  by  a  comparison 
of  food  and  feces,  but  attached  little  importance  to  the  observed 
increase  of  phosphorus  in  the  bones  of  the  lambs  which  had  received 
the  phosphates.      In  the  light  of  subsequent  investigation  in  this 


386 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


field  it  would  appear  that  the  increase  in  phosphorus  content  of  the 
bones,  as  caused  by  the  inorganic  phosphates,  was  sufficient 
definitely  to  increase  the  breaking  strength  of  the  bones.  The 
following  data  are  taken  from  Hof  meister's  tables. 

CALCIUM  AND   PHOSPHORUS   CONTENT  OF   BONES  OF   LAMBS  AS 
AFFECTED   BY  ADMINISTRATION  OF  BONE  PHOSPHATES 
Dry  Matter  Basis — Percent 


Jaw  bone 

Shoulder   blade 

Humerus 

Radius 

Tibia 

Ration 

P2O5 

CaO 

P2O5 

CaO 

P2O5 

CaO 

P2O5 

CaO 

P2O5 

CaO 

Low  phosphorus 

26.20 

31.30 

19.60 

24.50 

19.30 

22.60 

19.70 

23.90 

19.70 

24.30 

Low  phosphorus 
plus  bone  phos- 
phates 

27.80 

33.90 

20.50 

24.90 

19.40 

22.90 

19.90 

24.01 

20.30 

22.60 

Hofmeister  also  studied  the  digestibility  of  superphosphate 
(laboratory  prepared),  with  two  2-year-old  wethers,  and  concluded 
that,  with  small  doses,  all  the  water-soluble  lime  and  phosphoric  acid 
are  digested  from  superphosphate;  with  increased  doses,  one-third 
of  the  water-soluble  phosphoric  acid  is  digested,  while  all  the  water- 
soluble  and  half  of  the  insoluble  lime  is  digested.  The  facts  as  he 
noted  them,  would,  of  course,  be  differently  interpreted  at  this  time. 

Weiske  (1874)  notes  that,  of  animals  receiving  the  same  diet 
under  conditions  of  practice,  some  will  suffer  from  fragility  of  the 
bones,  and  others  will  not.  In  experiments  on  rabbits  receiving 
barley  and  various  mineral  supplements,  Weiske  found  that  the 

EFFECTS  OF  DIET  ON  THE  PHOSPHATES  OF  THE  BONES 
Weiske  (1874)  Percent,  Dry,  Fat-free  Basis 


Kind  of 
bone 

No.  of 
ani- 
mals 

Mineral 
matter 

P2O5 

CaO 

MgrO 

Feed 

Fowl, 
femur 
Fowl, 
femur 

1 
1 

45.64 
42.79 

16.67 
15.98 

23.14 
21.48 

0.64 
0.43 

Usual 
Same 

Normal.     Data  computed  by 

the  compilers. 
Showed  bone  disease. 

Lamb, 
pelvic 
Lamb, 
pelvic 
Lamb, 
pelvic 

1 

1 
1 

60.46 
60.36 
60.07 

23.94 
23.79 
23.94 

31.78 
31.60 
31.57 

Poor  in  phosphorus 

Poor  in  lime 

Normal 

These  are  bones    of  lambs  in 
experiments  reported  in  1873. 
Data  of  this  experiment  com- 
puted by  the  compilers. 

Rabbit 

Rabbit 
Rabbit 

2 

2 
2 

65.64 

65.08 
64.42 

26.79 

25.60 
26.06 

34.10 

33.68 
33.34 

0.75 

0.76 
0.76 

Calcium-free  barley; 

dist.  water 

Same  +  Mg--phosphate 

Same  +  Sr-phosphate 

Data  computed  by  compilers. 

Rabbit 

1 

65.62 

26.26 

34.23 

0.74 

Normal,  dist.  water 

5  mos.  old;  killed  when  feeding1 

began. 
6Vi  mos.  old;  killed  at  end  o 

experiment. 
7H  mos-  old. 

Rabbit 

Rabbit 
Rabbit 

1 

2 
2 

67.61 

69.03 
67.55 

26.87 

27.46 
26.95 

35.15 

35.92 
35.28 

0.80 

0.79 
0.73 

Normal,  dist.  water 

Normal,  dist.  water 
No  food;  dist.  water 

PHOSPHORUS  METABOLISM 


387 


percentage  content  of  bones,  in  ash  constituents,  varied  in  marked 
degree,  but  that  the  composition  of  the  bone  ash  remained  almost 
constant.  The  feeding  of  magnesium  phosphate  with  calcium-free 
food  did  not  cause  a  perceptible  increase  of  magnesium  in  the  bones, 
and  only  traces  of  strontium  were  found  in  the  ash  of  the  bones  of 
animals  to  which  strontium  phosphate  was  fed.  With  calcium 
lacking  in  the  food,  an  animal  lives  about  as  long  as  if  the  starvation 
is  total.  If  the  removal  of  the  mineral  matter  is  as  complete  as 
possible,  there  is,  as  in  total  starvation,  a  gradual  loss  of  bone  sub- 
stance. 

E.  Voit  (1880)  shows  that  lack  of  lime  in  the  diet  of  dogs 
reduces  not  only  the  lime  but  also  the  phosphorus  in  the  bones,  as 
set  forth  by  the  following  analyses : 

ANALYSES  OF  BONES  OF  YOUNG  DOGS  AS  AFFECTED  BY  LACK  OF 
LIME  IN  THE  FEED  (E.  Voit,  1880)— Percent 


Age    of 

Bone 

Part  of 
bone 

Water  con- 
tent of  skel- 
eton 

Percent  of  dry  substance 

pup 
Days 

Ash 

Pe203 

CaO 

Mg-O 

P2O5 

Treatment  of 
•     subject 

10 

Humerus 
Scapula 
Humerus 
Scapula 

Outer 
Outer 
Spongy 
Spong-y 

[■  66.2  -j 

32.25 
37.21 
30.45 
35.01 

0.06 
6.06 

17.04 
19.64 
15.92 
18.19 

0.39 
0.46 
0.40 
0.35 

12.93 
15.45 
12.25 
14.42 

/-  Killed  as  control 

38 

Humerus 
Scapula 
Humerus 
Scapula 

Outer 
Outer 
Spongy 
Spongy 

[■64.9-j 

41.32 
40.29 
33.71 
36.96 

0.18 
0.19 

6."3i 

21.40 
21.40 
16.87 
18.50 

0.38 

6.'  28 
0.33 

17.40 

12!  93 
13.62 

]  Fed  from  the  agre 
1  of  ten  days  for  28 
|  days,    lime    being- 
J  included 

38 

Humerus 
Scapula 
Humerus 
Scapula 

Outer 
Outer 
Spongy 
Spongy 

H 

30.94 
29.06 
26.40 
27.22 

0.16 
0.18 
0.16 
0.34 

16.36 
15.29 
13.34 
14.24 

0.32 
0.38 
0.21 
0.19 

13.17 

i6.*77 
10.21 

Fed  the   same   as 
[■last,  but    without 
lime 

J.  Cohnheim  (1882)  notes  that  Weiske  and  others,  and  Zalesky 
conclude  that  the  composition  of  the  ash  of  the  bones  is  not  affected 
by  the  composition  of  the  food,  but  that  Chossat,  Bibra,  Wegner, 
Voit  and  others  have  determined  that  the  bones  of  animals,  birds 
especially,  become  poor  in  earth  salts  and  easily  bent  if  the  food  is 
poor  in  these  constituents.  Young  lions  and  leopards  become 
rachitic  when  fed  on  meat  without  bones  (Roll,  1860) . 

Weiske,  Dehmel,  Kennepohl,  Schulze  and  Flechsig  (1885)  fed 
hay,  acidified  with  sulphuric  acid,  to  sheep,  from  Nov,  4,  1879  to 
Mar.  11, 1880,  and  studied  the  effects  of  the  acid  on  the  composition 
of  the  skeleton.  The  slight  differences,  indicating  a  solution  of  the 
bone  salts  by  the  acid,  were  regarded  by  the  authors  as  within  the 
legitimate  error  of  work.     A  portion  of  the  data  are  as  follows. 


•388  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

EFFECTS  OF  ACIDIFIED  HAY  ON  THE  COMPOSITION  OF  THE 
SKELETON  OF  SHEEP— Fat  and  Water  Free— Percent 


Lamb 
No. 

Feed 

Organic 
matter 

Mineral 
matter 

CaO 

MgrO 

CO2 

P2O5 

1 

38.16 
39.02 
39.96 

61.83 
60.98 
60.04 

31.55 
31.17 
30.34 

0.66 
0.63 
0.77 

3.07 
3.02 

24.26 

2 

23.88 

3 

Killed  at  beginning-  of  experiment 

23.59 

At  a  later  date  Weiske  (1886)  reported  more  data  on  this  same 
experiment,  and  showed  that  the  calcium  content  of  the  'flesh  was 
decreased  by  the  sulphuric  acid  feeding,  but  the  phosphorus  was  not 
altered ;  which  implies  that  the  effect  of  sulphuric  acid  on  the  bones 
in  the  experiment  of  Weiske,  Dehmel,  et  al.  (1885)  was,  at  least  in 
part,  due  to  its  interference  with  calcium  absorption. 

Weiske  (1888)  experimented  to  determine  the  effects  of  adding, 
during  6  months,  neutral  precipitated  calcium  phosphate  to  the  nor- 
mal food  of  rabbits.  The  phosphate  had  no  effect  on  the  gain  in 
weight,  nor  on  the  composition  of  the  bones,  though  there  was  a 
slight  increase  in  the  dry  fat-free  skeleton,  due  to  the  use  of  the 
phosphate. 

Henry  (1889, 1890)  reported  increase  in  strength  and  ash  of  the 
bones  of  corn-fed  pigs  by  the  administration  of  bone  meal.  Wood 
ashes  were  found  to  give  similar  though  less  prominent  results. 
Hard-water,  in  comparison  with  rain  water,  was  not  shown  to 
increase  the  ash  or  strength  of  the  bones.  No  increase  of  muscle 
was  found  (apparently  by  visual  examination)  to  result  from  the 
feeding  of  bone  meal  or  wood  ashes ;  nor  was  there  increase  in  the 
weight  of  the  blood  or  internal  organs. 

Beraz  (1890)  studied  the  teeth  of  dogs  as  affected  by  the  lime 
content  of  the  ration.  Rations  producing  rachitic  conditions  in  the 
skeleton  generally  did  not  produce  similar  effects  in  the  teeth. 

Weiske  (1891a)  quotes  earlier  work  showing  that  dilute  sul- 
phuric acid  would,  if  ingested  during  a  considerable  time,  lower  the 
calcium  content  of  both  bones  and  muscles,  and  refers  to  work  of 
Heitzmann,  Hofmeister  and  Siedamgrotzky  showing  that  if  lactic 
acid  be  given  the  specific  gravity  and  ash  content  of  the  bones  is 
subnormal,  and  to  work  of  Auerbach  showing  that  KH2P04, 
when  ingested  by  a  dog,  greatly  increased  the  ammonia  of  the  urine. 

Weiske  reports  work  of  his  own  in  which  NaH2P04  was  fed  to 
rabbits  on  a  ration  of  meadow  hay,  oats,  beets,  and  potatoes.  The 
analyses  of  the  skeletons  showed  no  differences  of  note  between  the 
rabbits  with  phosphate  and  those  without. 


PHOSPHORUS  METABOLISM 


389 


At  a  later  date  Weiske  (1891b)  reported  further  results  of  a 
similar  but  more  critical  study  with  rabbits,  the  rations  being  (1) 
hay,  (2)  oats,  (3)  hay  and  oats,  and  (4)  oats  and  NaH2P04.  From 
the  following  data  it  will  be  noted  that  the  hay  was  consistently 
superior,  as  a  bone  food,  to  hay  and  oats ;  that  the  hay  and  oats  were 
better  than  oats  alone;  and  that  the  rabbits  appeared  to  receive 
benefit  from  the  sodium  phosphate,  in  spite  of  its  acidity. 

COMPOSITION  OF  THE  BONES  OF  RABBITS  AS  AFFECTED  BY  THE 
FOOD  (Weiske,  1891b)— Percent,  Dry,  Fat-free  Bone 


Bones 


No.  of 

ani- 

Mineral 

P2O5 

CaO 

MgO 

mals 

matter 

1 

64.22 

24.89 

32.08 

0.77 

2 

63.78 

24.82 

v     31.83 

0.75 

2 

61.10 

23.90 

29.62 

1.01 

2 

61.59 

24.47 

30.25 

0.88 

1 

62.13 

23.73 

30.84 

0.68 

2 

61.55 

23.69 

30.66 

0.64 

2 

58.74 

22.69 

28.47 

0.92 

1 

58.47 

22.85 

28.55 

0.67 

1 

66.51 

26.02 

33.86 

0.73 

2 

66.65 

26.15 

33.62 

0.78 

2 

62.16 

24.85 

30.85 

0.94 

L 

63.38 

24.94 

31.76 

0.68 

1 

78.72 

33.88 

37.28 

2.63 

2 

77.93 

33.30 

36.73 

2.52 

2 

77.61 

33.36 

36.18 

2.75 

1 

80.01 

34.48 

38.08 

2.67 

Feed 


All 

All 

All 

All 

All  but  long  bones  and  teeth 
All  but  long  bones  and  teeth 
All  but  long  bones  and  teeth 
All  but  long  bones  and  teeth 

Long  bones 

Long  bones 

Long  bones 

Long  bones 

Teeth 

Teeth 

Teeth 

Teeth 


Hay 

Hay  and  oats 

Oats 

Oats  +Na  H2PO4 

Hay 

Hay  and  oats 

Oats 

Oats  +  Na  H2PO4 

Hay 

Hay  and  oats 

Oats 

Oats  +  Na  H2PO4 

Hay 

Hay  and  oats 

Oats 

Oats  +  Na  H2PO4 


Weiske  (1892)  reported  results  from  feeding  rabbits  on  oats, 
with  trisodium  phosphate,  monosodium  phosphate,  tricalcium  phos- 
phate, calcium  carbonate,  and  sodium  citrate  added  to  the  ration,  for 
different  lots  of  animals.  The  calcium  carbonate  greatly  improved 
the  oat  ration.  Acid  sodium  phosphate  made  the  oats  less  useful 
than  when  fed  alone,  and  the  effect  on  the  bones  was  injurious. 
Benefit  was  not  observed  from  the  other  salts. 

Arnstadt  (1893)  discusses  malnutrition  of  the  bones  of  cattle 
and  other  farm  animals  as  caused  by  lack  of  phosphorus  in  the  food. 
Pregnant,  milk-producing,  or  young,  growing  animals  were  found  to 
suffer  most  severely.  He  speaks  of  liberal  allowance  of  common  salt 
being  of  benefit  when  fed  with  calcium  phosphate,  through  assisting 
in  its  assimilation. 

Weiske  (1895a)  fed  oats,  having  an  acid  ash,  to  rabbits,  with  and 
without  other  food,  except  for  calcium  sulphate  and  tricalcic  phos- 
phate, which  were  fed  to  certain  individuals.  There  was  a  loss  of 
mineral  matter  from  the  bones,  but  no  great  change  in  composition. 
The  sulphate  and  phosphate  of  calcium  proved  not  to  be  so  valuable 
as  previous  experiments  had  shown  the  carbonate  to  be,  when  added 
to  the  same  ration.     . 


390 


OHIO  EXPEEIMENT  STATION:  TECHNICAL  BUL.  5 


Weiske  (1895c)  reports  results  of  feeding  experiments  on  cal- 
cium-poor rations,  which  were  rich  in  phosphorus,  and  to  which  were 
added  as  supplements,  calcium,  strontium  and  magnesium  carbon- 
ates, and  calcium  sulphate.  The  alkaline  earth  carbonates  were 
found  of  value  in  the  correction  of  the  acidity  of  the  ash  of  an  oat 
ration,  but  magnesium  and  strontium  do  not  physiologically  replace 
calcium  in  the  skeleton,  although  not  inconsiderable  quantities  of 
these  elements  were  transferred  there.  The  urine  of  the  rabbits 
receiving  the  alkaline  earths  was  turbid  and  became  continually 
more  alkaline.  The  rabbit  on  oats  alone  had  an  acid  urine.  The 
addition  of  calcium  carbonate  was  much  more  beneficial  in  its  effect 
on  the  gain  in  weight  than  were  other  mineral  supplements. 

COMPOSITION  OF  THE  BONES  OF  RABBITS,  AS  AFFECTED  BY  THE 
FOOD   (Weiske,  1895c)— Percent,  Dry,  Fat-free  Bone 


Bones 


No.  of 

ani- 

Mineral 

P2O5 

CaO 

MgO 

mals 

matter 

61.34 

23.42 

31.43 

0.73 

60.17 

23.76 

30.72 

1.11 

55.47 

22.21 

23.02 

0.61 

56.98 

23.21 

28.36 

1.39 

56.62 

22.69 

28.85 

0.63 

67.14 

25.75 

34.70 

0.85 

64.58 

25.78 

32.98 

0.76 

62.59 

25.76 

27.95 

0.96 

63.89 

25.48 

32.23 

1.48 

61.73 

24.44 

31.63 

0.74 

78.66 

33.63 

38.44 

2.40 

79.35 

34.71 

38.74 

2.61 

77.58 

34.45 

36.96 

3.23 

77.74 

33.37 

37.72 

2.57 

Feed 


All  but  loner  bones  and  teeth 
All  but  long:  bones  and  teeth 
All  but  loug-  bones  and  teeth 
All  but  long  bones  and  teeth 
All  but  long-  bones  and  teeth 

Long-  bones 

Long-  bones 

Long-  bones 

Long-  bones 

Long-  bones 

Teeth 

Teeth 

Teeth 

Teeth 


Oats+  CaC03 

O  ats  +Ca  SO4 

Oats  +  Sr  CO3 

Oats  +  MgC03 

Oats 

Oats  +  Ca  CO3 

Oats  +  Ca  SO4 

Oats  +  Sr  CO3 

Oats  +  Mg-C03 

Oats 

Oats  +  Ca  COs 

Oats  +  Ca  SO4 

Oats  -+•  Mg-C03 

Oats 


Springer  (1902)  reports  that  by  the  application  of  electricity 
for  5  minutes  daily  at  the  seat  of  the  epiphyseal  cartilages  of  the 
legs,  with  rabbits,  he  induced  an  added  growth  of  the  bones  as  indi- 
cated by  the  following  figures : 


Weight  of  Femur 


Length  of  Femur 


Grams 

Treated  1     1.105 


Meters 

0.066 

1.185     0.064 


Controls  3     0.977 


0.060 


0.940     0.059 


Aron  (1905)  states,  with  reference  to  bone  diseases  of  animals, 
as  caused  by  abnormal  food,  that  with  high  potassium  and  low 
sodium  intake,  bone  formation  is  held  below  normal,  even  though 
sufficient  lime  and  phosphorus  be  taken,  potassium  chloride  in  solu- 
tion with  primary  phosphates  apparently  tending  to  prevent  their 
change  to  di-  and  tri-calcic  salts. 


PHOSPHORUS  METABOLISM  391 

Rasquin  (1905-6)  fed  two  young  cocks,  one  with  and  one  with- 
out powdered  bone,  for  120  days.  Under  the  influence  of  these 
rations  the  fowls  developed  as  indicated  by  the  following  data: 

A  (With  Bone)         B  (Without  Bone) 

Weight  of  body,  live 2.690  kg 2.470   kg. 

Weight   of   body,   plucked 2.455  kg 2.240  kg. 

Weight  of  body,  drawn 2.003  kg 1.790  kg. 

Weight  of  skeleton 236  gm 190  gm. 

Weight  of  ischium 16  gm 13  gm. 

Weight  of  bones  of  wings   41  gm 30  gm. 

Weight  of  bones  of  legs 83  gm 58  gm. 

Weight  of  breast  bone 12  gm '.      8  gm. 

Lime  in  bones   21.98  pet 20.51  pet. 

Phosphoric  acid  in  bones   18.77  pet 18.14  pet. 

Length  of  branches  of  breast  bone 13   cm 9  cm. 

Burnett  (1906,  1908,  1910)  examined  the  bones  of  hogs  which 
had  received  natural  foods  varying  in  calcium  and  phosphorus  con- 
tent, and  also  others  which  had  received  with  the  food  bone  meal, 
calcium  carbonate,  or  disodium  phosphate.  The  development  of  the 
bones,  as  determined  by  breaking  strength,  specific  gravity,  thick- 
ness of  wall,  ash  content,  etc.,  reflected  the  character  of  the  food  in 
the  usual  way.  The  principal  deposit  of  bone  salts  seems  to  have 
been  inside  the  marrow  space,  the  walls  in  this  way  becoming 
greatly  thickened.  Summary  tables  from  three  series  of  observa- 
tions are  quoted  on  pages  392  and  393. 

These  tables  from  the  experiments  of  Burnett  show,  more 
plainly  than  any  of  the  earlier  work,  the  fact  that  the  mineral  con- 
tent of  the  bones  may  be  modified,  in  accordance  with  the  composi- 
tion of  the  foods,  through  a  very  wide  range  of  variation.  If  this 
fact  appears  more  prominent  in  relation  to  swine  than  to  other  ani- 
mals it  is  doubtless  due  to  the  unusual  mineral  content  of  some  of 
the  rations  used  for  swine.  Through  the  feeding  of  precipitated 
bone  flour,  which  is  readily  taken  by  the  larger  farm  animals,  it  is 
probably  possible  to  affect  the  mineral  content  of  their  bones  also 
to  such  extent  as  to  be  of  practical  importance,  especially  with 
horses.      See  also  the  experiments  of  Forbes  (1909)  p.  374. 

Alway  and  Hadlock  (1909)  compared  hog  bones  produced  from 
corn  alone  with  others  produced  from  corn  and  bone  meal.  They 
state  that  both  lots  had  practically  the  same  composition,  thus  indi- 
cating that  the  nature  of  the  food  has  no  effect  upon  the  relative 
proportions  of  the  different  constituents  of  the  mineral  matter  of 


392 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


bones.  At  the  same  time,  considering  the  bones  from  corn  and  bone 
meal  as  the  standard,  the  feeding  of  corn  alone  is  shown,  by  their 
ash  analyses,  to  have  increased  the  magnesium  by  20  percent,  to 
have  increased  the  phosphorus  1.7  percent  and  to  have  decreased 
the  calcium  by  1.0  percent  of  the  amounts  present.  A  part  of  their 
figures  are  as  follows: 

Weak  bones         Strong  bones 

(Corn)  (Corn;  bone  meal) 

Average  percent  mineral  matter,  green  bones  25.4 37.2 

Average  wall  thickness,  green  bones 2.8  mm 4.1  mm. 

Average  breaking  strength,  green  bones  ....702.0   lbs 1505.0  lbs. 

CaO  52.47  percent       52.98  percent 

MgO 1.24  percent         1.05  percent 

P205 , 41.61  percent       40.93  percent 

EFFECTS  OF  FOODS  ON  THE  BREAKING  STRENGTH  OF  THE  BONES 
OF  SWINE  (Burnett,  1906) 

FIRST  TEST 


Killed  Feb.  10- 

Number  in  lot 

Ration 

Av.  live  weight,  lbs. . 
Breaking  strength  of 

radius,  lbs 

Breaking  strength  of 

tibia,  lbs 

Av.  of  two  bones  in 

each  leg,  lbs 


Lot  I 


Soaked  corn 
192 
723 
607 
714 


Lot  II 


Corn  90% 

Tankage  10% 

247 

1308 

825 

1007 


Lot  III 


3 

Corn  90% 

Ground  bone  10  % 

"257 

1561 

732 
1081 


Lot  IV 


Corn  75% 

Shorts  25% 

210 


641 

783 


SECOND  TEST 


Killed  March  14— 

4 
171 

768 

542   * 

634 

4 

286 

1254 
834 
1057 

4 
242 

1044 

779 

942 

4 

Av.  live  weight,  lbs. . . 
Breaking  strength  of 

204 
835 

Breaking  strength  of 

662 

Av.  of  two  bones  in 

801 

AVERAGE  BREAKING  STRENGTH  OF  BONES  PER  100  POUNDS  LIVE 
WEIGHT  OF  HOGS  AT  TIME  OF  SLAUGHTER  (Burnett,  1908)— Pounds 


Let 

Ration 

Femur 

Tibia 

Humerus 

Radius 
and  ulna 

All  bones 

1 

276 
343 
462 
559 
646 

252 
309 
360 
409 
465 

434 

555 
685 
740 
898 

341 

376 
529 
611 
715 

325 

?, 

396 

3 

509 

4 

580 

5 

681 

PHOSPHORUS  METABOLISM 


393 


rf  (« r  g 

cciomojco 

C  M  S-O 

OOCOWCOIO 

Perce: 
mini 
matt 

green 

CO  CO  CO  "-* ''Jl 

<o       S  01    • 

b0_,  i  «S 
rt—  Q  G  _ 

cd  oo  t-  t-iTH 

Aver 

wa 

thick: 

of  bo 

in  n 

■^i^-^llOiO 

rage 
;ific 
vity 
ones 

Hr-tMr-IM 

ococococo 

CO  CM  CM  CO  CO 

v  5  dfi 

>  nhH 

«3  ol  e*  o 

H 

M 

SGcu 

H 

irii— i<ooai 

Aver; 
volui 
of  bo 
in  c. 

IOCDtHo'^) 

O 

w 

COHOHH 
i-Hi-Hi-trH 

Average 
weight 
of  bones 
in  gms. 

co  co  ^a>  co 

CM  ••31  CO  ■*  "0 

& 

I-H  T-H  I— 1 1— 1  rH 

w 

« 

H 

, 

o 

<5   . 

CO 

o 

CD 

r-l 

00 

Average 

circumfer 

ence  of 

bones  in 

mm. 

CO  CO  CC  CM  CO 

cDt-cot-c-" 

o 

CO 
H 

co 

«*^ 

,   -+* 

03 

CO 

W  2 
H  g 

Average 

length  ol 

bones  in 

mm. 

C5t-CNr-CO 

rH 

Fh 

o  S 

O 

m 

w 

H 

CD  CO  CD  CO  CD 

O 

OS 

3h 

u 

Average 
breaking 
trength  of 
bones  per 

100  lbs. 
ive  weight 

of  hog 

fc 

Ph 

10C001I-* 

« 

O 

cocnt-cnr- 

■WCOTfl'*10 

o 

H  ^ 

!x 

s* 

01                -* 

Pn 

©  w 

CO 

Is 

A  vera 
breaki 
streng 
of  bon> 
.     in  lbs 

CCOW«H 

p 

02 

p 

CO  CO  CM  lO  CO 

r— t-oocncci 

5?    5    ** 
2  £  w    c 

■^xcpmco.— i 

coa5t-c»t— 

^1    1 

i-H  r-t  t— <  r-t  rH 

fe 

O 

CQ 

H 

U 

H 

fe 

Ph 

H 

R 

o3  i 

a 

o  a  c 

J3  3  s 

■4J 

01  ££ 

ri 

« 

J -a  "St 

scfic 

ai  rt  rt  c 

I 

S  —  —  - 

rt  is  o 

s  ii  :'  : 

^SEE 

S3  rt  rt  c 

,  (sssas 

_.  rt  rt  c 

! 

,T3  **  a  N 

:  ^"3  rt" 

i 

esses 

o  o  o  o  < 

i 

c 

c 

>c 

>c 

>c 

> 

OOHOOrt 

OCM1T305-* 
CO  CO  CO  CO  T»f 


cm  cor- CD  CO 

CMoicMCO'^5 


COt— CDOCO 
■**  CO  ~H  C<1  CO 
Hi- 1  CM  CO  CO 


CO  CD  CD  00  CD 


i— IOCOCOO 

•hi  cm  co  coco 

OCMCMCO'"* 


CD  CD  CD  COt- 


COCO  en  cm  in 
cd  co  cor— co 


ioin<— ICDCM 
r-t-ot--H 

CN  CM  CO  CM-* 


■^ICOt—  i-HlO 
r—  CD  COt—  05 
COCOTJICDr— 


CD'-nmcoco 
co^m^co 


«  a  £ 

.gSfl 
:a  oi+j^i 

•'d'O'O'ci 
a  a  a  a 

pj  cd  ra  rt 
ddddd 

O  O  O  O  o 

ooooo 


•s  a 
p.  a 


394  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Hart,  McCollum  and  Humphrey  (1909),  in  their  study  of 
phytin  metabolism  with  a  milch  cow,  found  that  when  calcium  or 
phosphorus  was  deficient  in  the  food,  the  skeletal  tissues  appeared  to 
be  ready  sources  of  supply.  The  average  quantities  of  calcium 
oxide  and  phosphorus  pentoxide  metabolized  and  excreted  daily 
during  periods  of  deficient  supply  were,  respectively,  50  and  60  gm. 

Schaumann  (1910)  says,  "The  idea  that  the  phosphorus  elimin- 
ated in  the  urine  in  pronounced  phosphorus  starvation  comes  mainly 
from  bony  tissue,  as  Cathcart  thinks,  is  supported  by  the  findings 
of  Hoist  with  guinea  pigs,  of  Lipschutz  and  of  Heubner  with  dogs, 
and  by  my  own  with  rabbits."  (Lipschutz,  1910a;  Heubner,  1909; 
Schaumann,  1908.) 

Lipschutz  (1910a)  studied  phosphorus  metabolism  in  dogs  with 
rations  varying  in  phosphorus  content.  A  ration  very  low  in  phos- 
phorus produced  a  moderate  increase  in  weight,  but  at  the  end  of 
seven  weeks  there  were  protracted  muscular  cramps  and  other 
symptoms  of  general  physiological  derangement.  An  examination 
of  the  bones  showed  a  condition  which  he  describes  in  detail  as 
resembling  Barlow's  disease.  A  ration  containing  the  same  amount 
of  calcium,  but  more  phosphorus,  caused  normal  development — a 
case  of  phosphorus  limiting  the  utilization  of  calcium.  Shortage  of 
calcium  limiting  the  use  of  phosphorus  is  much  more  common  in  the 
practical  feeding  of  animals. 

R.  Berg  and  C.  Rose  (1910)  state  that  children  and  soldiers 
from  regions  of  hard-water  have  fewer  unsound  teeth  than  others 
from  soft-water  regions.  The  facts  may  be  as  stated,  but  are  diffi- 
cult of  satisfactory  proof,  especially  as  due  to  the  water. 

Dibbelt  (1911)  reported  on  experiments  on  the  production  and 
cure  of  experimental  osteomalacia.  For  several  months  a  dog  was 
kept  on  a  low-calcium  diet  to  which  were  added  disodium  phosphate 
and  sodium  chloride.  At  the  end  of  this  time  the  left  fourth  rib 
was  removed  for  analysis.  Then  after  12  days,  in  which  dicalcium 
phosphate  replaced  the  sodium  phosphate,  the  right  fourth  rib  was 
removed.  For  the  next  3  months  the  dog  was  kept  on  a  mixed 
diet  with  liberal  allowance  of  bones.  At  the  end  of  this  period  the 
left  fifth  rib  was  removed.  During  the  experiment  the  dog  gave 
birth  to  four  young.  The  analysis  of  the  ribs  showed  that  the  with- 
drawal of  calcium,  as  a  result  of  the  diet  and  the  bearing  of  young, 
caused  an  atrophy  of  the  bone  tissue,  and  that  the  administration  of 
the  calcium  salt,  and  omission  of  the  salts  causing  the  withdrawal 
of  calcium,  with  normal  feeding,  had  the  effect  to  replace  this  loss. 


PHOSPHORUS  METABOLISM  '395 

For  earlier  views  on  calcification  of  bone  see  Aeby  (1873b).  For 
other  material  on  the  effects  of  diet  on  the  composition  of  the  bones 
seeMalyand  Donath  (1873),  Weiske  (1873,  1891b),  Laurer 
(1910),  and  Cagnetto  (1911) ;  also  Hart,  McCollum  and  Fuller 
(1909),  p.  328  of  this  work,  and  E.  B.  Forbes  and  associates  (1914). 

Summary  on  Common  Foods  in  Relation  to  Phosphorus  Metabolism 

Our  knowledge  of  the  specific  effects  of  common  foods  in  rela- 
tion to  phosphorus  metabolism  is  slight  and  fragmentary.  The 
evidence,  however,  is  of  such  nature  as  to  promise  important  results 
from  further  study. 

The  retention  of  phosphorus  from  common  foods  is,  in  general, 
in  the  same  order  as  the  intake,  but  deficiency  of  nutrients  (as 
calcium)  necessary  to  the  maximum  utilization  of  phosphorus 
may  limit  retention,  as  also  may  excessive  amounts  of  magnesium 
in  the  ration.  The  deposit  of  calcium  phosphate  in  the  bones  is 
also  hindered  by  inorganic  acids,  acid  salts,  calcium-precipitating 
ions  other  than  phosphorus,  and  a  high  potassium  and  low  sodium 
content  of  the  ration. 

The  human  being  stores  more  phosphorus  from  whole  wheat 
bread  than  from  white  bread. 

In  comparing  diets  containing  meat  and  fish,  the  latter 
appears  more  favorable  to  phosphorus  and  magnesium  retention 
but  less  favorable  to  calcium  retention  than  the  former. 

White  rice  does  not  contain  as  much  phosphorus  as  the  adult 
human  being  requires.  Red  or  unpolished  rice  contains  more  than 
enough. 

Beet  molasses  has  been  thought  to  increase  the  solubility  and 
therefore  the  assimilability  of  the  phosphorus  of  the  ration  of  the 
horse. 

Young  carnivora  suffer  from  malnutrition  of  the  bones  if  fed 
on  meat  alone,  to  the  exclusion  of  bone. 

The  low  calcium  content  of  cereals  and  other  seeds  has  been 
shown,  in  experiments  with  growing  swine,  to  limit  the  retention  of 
phosphorus.  The  addition  of  calcium  to  rations  composed  of 
cereal  foods  serves  to  increase  phosphorus  retention. 

In  experiments  with  growing  swine,  comparing  many  common 
food  during  the  feeding  of  skim  milk  there  were  minimum  propor- 
tions of  potassium,  magnesium,  sulphur,  chlorine  and  phosphorus  in 
the  feces,  and  maximum  retention  of  calcium,  magnesium,  sulphur 
and  phosphorus. 


896  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Experiments  with  growing  swine  show  that  many  common 
foods  are  lacking  in  the  amount  of  calcium  and  phosphorus  neces- 
sary to  maximum  production  of  tissues  containing  phosphorus. 

Foods  have  been  shown  to  affect  the  phosphorus  content  of  the 
muscle,  the  liver  and  the  kidneys  of  swine,  as  related  to  the  ash,  the 
nitrogen  and  the  fresh  substance  of  these  parts.  The  total  and 
inorganic  phosphorus  of  the  blood,  the  lecithin  content  of  the  liver, 
and  the  composition  of  the  ash  of  the  bones  of  pigs  have  also  been 
shown  to  be  susceptible  of  variation  through  the  influence  of  corn  in 
the  diet. 

The  bones  may  be  greatly  weakened  by  protracted  feeding  on 
foods  poor  in  either  calcium  or  phosphorus,  and  may  be  greatly 
strengthened  by  the  use  of  foods  rich  in  these  elements,  the  bones 
appearing  to  possess  an  extensive  storage  function  for  calcium, 
magnesium  and  phosphorus. 

Rations  which,  because  of  deficiency  of  mineral  salts,  cause 
rachitic  changes  in  the  skeleton,  have  not  been  shown  to  produce 
such  effects  on  the  teeth,  though  it  has  been  stated,  with  some 
evidence  as  a  basis,  that  the  salts  of  the  drinking  water,  in  the 
course  of  time,  may  affect  the  quality  of  the  teeth. 

In  the  effects  of  diet  on  the  composition  of  the  bones,  the  ratio 
of  the  mineral  constituents,  one  to  another,  varies  but  little ;  there 
is  little  replacement  possible,  for  instance,  of  calcium  phosphate  by 
strontium  phosphate,  though  there  is  some  change  of  proportion  of 
calcium,  magnesium  and  phosphorus  in  response  to  the  character  of 
the  food.  The  total  ash  content  of  the  bone  may  be  varied  almost 
at  will,  by  the  administration  of  calcium  phosphate  or  by  the  use  of 
foods  containing  the  same  elements. 

Lack  of  food  also  affects  the  phosphorus  content  of  the  bones, 
since  these  tissues,  along  with  the  soft  parts,  undergo  katabolism  in 
starvation. 

Calcium  phosphate  in  the  diet  does  not  greatly  influence  gain  in 
body  weight,  but  is  readily  deposited  in  the  bones,  especially  within 
the  marrow  spaces,  having  the  effect  to  increase  the  density  and 
strength  of  these  supporting  structures. 

The  variations  in  phosphorus  content  of  milk,  ascribable  to 
effects  of  food,  are  within  the  range  of  normal  variation. 


PHOSPHOKUS  METABOLISM  397 

PHOSPHORUS  REQUIREMENTS  OF  ANIMALS 

GENERAL  DISCUSSION 

In  the  practical  nutrition  of  human  beings,  as  well  as  live  stock, 
our  interest  is  especially  in  conditions  of  maximum  production  or 
efficiency.  We  never  feed  for  mere  maintenance.  Maintenance 
expense  is  all  loss.  Our  interest  is  in  production,  and  the  amounts 
and  kinds  of  nutriment  necessary  to  sustain  maximum  production 
can  with  certainty  be  determined  only  by  production. 

Optimum  conditions,  however,  are  indefinite  and  variable.  We 
know  almost  nothing  as  to  the  maximum  amounts  of  phosphorus 
useful  to  animals  in  their  various  states  of  life  and  activity,  and  are 
quite  unable  to  submit  accurate  figures  as  guides  to  practice. 
Maintenance  requirements  are  more  accurately  known,  are  less 
variable  with  conditions,  and  hence  may  be  more  definitely 
expressed  with  safety;  but  as  guides  in  practical  nutrition  they 
must  be  regarded  as  irreducible  minima,  and  hence  as  danger 
signals,  and  distinctly  not  as  expressions  of  the  amounts  which 
animals  should  receive.  For  real  guides  to  practice  we  must,  for 
the  present,  at  least,  depend  in  large  measure  on  very  general 
recommendations,  and  on  that  judgment  which  is  the  result  of 
experience. 

Our  opinions  as  to  phosphorus  requirements  of  animals  are 
based  principally  on  balances  of  intake  and  outgo,  and  data  on 
elimination  during  fast.  In  the  use  of  such  observations  we  must 
bear  in  mind,  first,  that  a  given  amount  either  of  gain  or  loss  of 
phosphorus  in  the  body  ordinarily  represents  a  larger  amount  of 
food  phosphorus.  If  an  animal  gains  one  gram  per  day  of  phos- 
phorus on  an  intake  of  3  grams  it  would  not  do  to  assume  that  2 
grams  represents  the  maintenance  requirement.  The  one  gram  of 
phosphorus  retained  by  the  animal  represents  more  than  one  gram 
of  food  phosphorus,  because  there  are  certain  losses  inevitably 
incident  to  its  utilization,  and  there  would,  therefore,  be  a  certain 
retention  of  phosphorus,  with  the  above  mentioned  animal,  on  a  2.Q 
gram  intake;  second,  a  part  of  the  phosphorus  katabolized  by  the 
tissues  and  excreted  into  the  intestine  is  normally  reabsorbed  and 
utilized,  as  is  shown  by  the  fact  that  the  phosphorus  loss  from  the 
body  is  less  on  a  phosphorus-free  diet  than  during  fast,  the  reason 
for  which  is  not  definitely  known,  but  which  is  perhaps  connected 
with  the  greater  absorptive  activity  in  the  intestine  during  the 
digestion  of  food  than  during  fast,  a  fact  which  would  make  the 
fasting  loss  greater  than  the  minimum  maintenance  requirement, 
were  it  not  that  this  factor  is  offset  by  the  compensating  considera- 
tion that  a  part  of  the  food  phosphorus  normally  escapes  digestion. 


398  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

As  a  matter  of  fact  these  two  opposing  factors  must  commonly 
somewhat  nearly  balance  each  other  in  human  nutrition,  for  the 
minimum  maintenance  requirement  and  the  fasting  loss  happen  to 
be  so  nearly  the  same  that  the  one  may  be  used  as  a  measure  of  the 
other  without  great  error.  This  is  not  necessarily  so  in  animal 
nutrition  generally,  however,  for  were  the  maintenance  require- 
ment determined  with  foods  containing  phosphorus  in  indigestible 
relations  to  other  nutrients,  the  loss  of  indigestible  food-phosphorus 
might  exceed  the  reabsorption  of  once-digested  and  excreted  phos- 
phorus from  the  intestine,  so  that  the  fasting  loss  would  not  equal 
the  maintenance  requirement.  The  phosphorus  maintenance 
requirement  of  a  horse,  for  instance,  would  be  less  on  fresh  grass 
than  on  hay  made  from  the  same  grass,  because  of  the  greater 
apparent  digestibility  of  the  phosphorus  of  the  former  than  of  the 
latter. 

Since  it  is  impossible  to  feed  an  animal  so  as  to  maintain  it  in 
exact  phosphorus  equilibrium  we  are  obliged  to  form  our  judgments 
as  to  requirements  from  data  on  experiments  in  which  the  subject 
either  gained  or  lost  phosphorus.  From  such  data  we  are  unable  to 
state  the  phosphorus  requirement  in  an  exact  way,  even  for  the  con- 
ditions of  the  experiment,  since,  as  above  stated,  either  gain  or  loss 
of  phosphorus  represents  more  than  the  same  amount  of  phospho- 
rus in  the  food,  under  ordinary  conditions  of  life.  A  gram  of  phos- 
phorus retention,  however,  represents  the  same  amount  of  food 
phosphorus  as  a  gram  of  loss  of  phosphorus  from  the  body,  and 
hence  by  subtracting  the  retention  of  phosphorus  from  the  amount 
in  the  food,  or  adding  the  amount  of  the  loss  of  phosphorus  to  the 
amount  in  the  food,  we  are  able  to  say  that  the  requirement  is  less 
than  the  former,  and  greater  than  the  latter.  The  smaller  the 
retention  or  loss  the  more  closely  will  the  apparent  requirement, 
calculated  as  above,  approximate  the  actual  requirement  under  the 
conditions  of  the  determination. 

In  order  accurately  to  determine  the  phosphorus  requirements 
of  animals  the  ration  should  be  so  compounded  that  deficiencies  of 
nutrients  other  than  phosphorus,  but  essential  to  the  formation  of 
the  phosphorus  compounds  of  the  body,  shall  not  limit  the  useful- 
ness of  the  phosphorus  of  the  food.  These  conditions  are  not 
always  observed  in  studies  of  this  sort,  and  to  this  is  due  a  large 
measure  of  the  variability  of  the  results.  It  is  true  that  a 
considerable  measure  of  independence  of  other  nutrients  is  to  be 
seen  in  the  retention  of  phosphorus,  but  at  the  same  time  nitrogen 
and  calcium,  especially,  are  so  largely  used  with  phosphorus  in  the 
body  that  coincident  negative  balances  of  both  nitrogen  and  calcium 
would  in  a  short  time  come  to  exercise  a  retarding  influence  on  the 
retention  of  phosphorus. 


PHOSPHOKUS  METABOLISM 


399 


Other  factors  entering  into  the  determination  of  results  of 
balance  experiments  with  animals  are  the  food  habit,  as  determined 
by  previous  feeding,  the  nature  of  the  particular  foods  used,  and 
also  the  kind,  age  and  weight  of  the  subject. 

From  among  the  many  balance  experiments  which  we  might 
use  in  this  discussion  of  phosphorus  requirements  we  present  a  few 
typical  reports  of  each  of  the  various  sorts,  without  an  effort  to  use 
them  all.  On  account  of  the  diversity  of  conditions  attending  these 
experiments  the  computation  of  mathematical  averages  would 
constitute  an  act  of  scientific  violence  of  which  we  desire  not  to  be 
convicted. 

The  reader  will  probably  be  surprised  to  note  the  extremely 
fragmentary  character  of  the  data  even  on  minimum  phosphorus 
requirements  of  animals,  while  on  optimum  requirements  we  have 
almost  no  evidence  at  all.  Most  writers,  thus  far,  have  been 
obliged  to  submit  computed  estimates  only,  and  as  yet  there  is  but 
slight  basis  for  more  definite  statements. 

PHOSPHORUS  EXCRETION  DURING  FAST 

In  studying  phosphorus  excretion  during  fasting  Ajello  and 
Solaro  (1893)  found,  with  Giovanni  Succi  as  a  subject,  that  the 
phosphorus  excretion  in  the  urine  varied  as  the  loss  in  body  weight, 
and  not  as  the  volume  of  the  urine. 

C.  Lehmann,  et  al.  (1893)  showed  that  the  average  daily  excre- 
tion of  P205  in  10  days  fasting  with  Cetti  (urine  and  feces  both 
included)  was  2.675  gm. ;  with  Breithaupt  the  average  daily  P205 
excretion  during  6  days  was  2.28  gm. 

E.  and  0.  Freund  (1901)  investigated  the  proportions  of  acid, 
alkaline  and  earthy  phosphates  in  the  urine  during  fast.  The  follow- 
ing figures  show  the  results : 


FOI 

IMS  OF  PHOSPHORUS 

IN  URINE 

DURING  FAST— Grams 

P*Oe 

Day  of 

Earthy 

Acid 

Alkaline 

Day 

Earthy 

Acid 

Alkaline 

Day 

Earthy 

Acid 

Alkaline 

phos- 

phos- 

phos- 

of 

phos- 

phos- 

phos- 

Of 

phos- 

phos- 

phos-   j 

phates 

phates 

phates 

fast 

phates 

phates 

phates 

fast 

phates 

phates 

phates 

1 

0.19 

1.60 

1.38 

8 

0.47 

1.08 

1.23 

15 

0.46 

0.37 

0.95 

2 

0.26 

1.41 

1.34 

9 

0.81 

0.85 

1.55 

16 

0.36 

0.25 

0.63 

3 

0.48 

1.40 

1.12 

10 

0.48 

0.54 

1.14 

17 

0.39 

0.38 

0.96 

4 

0.21 

1.48 

1.06 

11 

0.30 

0.45 

0.96 

18 

0.32 

0.27 

0.69 

5 

0.27 

1.46 

1.05 

12 

0.51 

0.46 

0.89 

19 

0.33 

0.39 

0.74 

6 

0.45 

13 

0.35 

0.36 

0.68 

20 

0.17 

0.26 

0.41 

7 

0.39 

0.91 

1.22 

14 

0.41 

0.29 

0.70 

21 

0.14 

0.23 

0.41 

Ave. 

7  days 

0.32 

1.21 

1.19 

0.48 

0.58 

1.02 

0.31 

0.31 

0.67 

From  these  data  it  is  apparent  that,  considering  the  results 
from  a  week  together,  there  was  during  this  fast  a  progressive 
decrease  in  both  acid  and  alkaline  phosphates,  while  the  earthy 


400 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


phosphates  were  the  same  in  amount  in  the  last  week  as  in  the  first. 
Averaging  these  weekly  figures  shows  that  for  the  whole  fast  the 
daily  excretion  of  earthy  phosphates  was  0.37  gm.,  of  acid  phos- 
phates 0.70  gm.,  and  of  alkaline  phosphates  0.96  gm.,  stated  as  P205. 
Gilbert  and  Posternak  (1903)  state  that  the  loss  of  phosphorus 
in  the  urine  of  fasting  subjects  is  as  follows. 
DAILY  ELIMINATION  OF  P205  IN  URINE— Grams  per  Kilo  Body  Weight 

Man    0.035 

Dog    0.065 

Cat     0.110 

Cathcart  and  Fawsitt   (1907)   have  given  us  the  following 
figures  on  fasting  metabolism. 

DAILY   URINARY   EXCRETION   OF   INORGANIC   ELEMENTS   DURING 

FAST 


S 

S 

S 

•Acidity 

Day  of 

P2O5 

CI 

(total) 

(inorganic) 

(neutral) 

Exp. 

C.C  N/10 

Diet 

Grams 

Grams 

Grams 

Grams 

Grams 

NaOH 

4 

3.60 

7.3 

1.39 

1.17 

0.14 

579 

Egg  and  milk 

5 

3.73 

5.4 

1.41 

1.13 

0.18 

588 

Egg  and  milk 

6 

4.14 

6.7 

1.33 

1.07 

0.15 

582 

Egg-  and  milk 

1 

2.26 

3.2 

0.62 

0.44 

0.13 

378 

Past 

2 

2.93 

2.0 

0.93 

0.75 

0.12 

640 

Fast 

3 

2.98 

1.5 

0.80 

0.65 

0.12 

687 

Fast 

4 

2.91 

1.3 

0.86 

0.69 

0.12 

604 

Fast 

6 

2.37 

1.0 

0.71 

0.55 

0.12 

454 

Fast 

7 

1.84 

0.84 

0.64 

0.50 

0.11 

358 

Fast 

8 

1.89 

0.59 

0.62 

0.49 

0.09 

344 

Fast 

10 

1.60 

0.39 

0.56 

0.43 

0.10 

280 

Fast 

11 

1.54 

0.30 

0.57 

0.42 

0.12 

256 

Fast 

12 

1.55 

0.18 

0.58 

0.44 

0.11 

212 

Fast 

14 

1.25 

0.24 

0.54 

0.39 

0.11 

228 

Fast 

1 

0.45 

0.53 

0.48 

0.31 

0.12 

108 

Starch  and  cream 

2 

0.20 

0.66 

0.28 

0.14 

0.10 

88 

Starch  and  cream 

3 

0.34 

0.97 

0.29 

0.15 

0.10 

104 

Starch  and  cream 

4 

0.89 

1.10 

0.93 

0.73 

0.16 

156 

Egg  and  milk 

5 

2.10 

4.60 

0.93 

0.74 

0.16 

263 

Egg  and  milk 

Thus  the  urinary  phosphorus  excretion  during  fast  varies 
between  2.98  and  1.25  gm.  P205,  the  average  being  2.10  gm.  daily. 
The  decreased  excretion  of  P205  after  the  end  of  the  fast  is  notable, 
and  is  coincident  with  a  similar  fluctuation  in  the  excretion  of 
sulphur.      It  doubtless  signifies  synthesis.- 

Benedict  (1907),  in  connection  with  his  own  work  on  fasting 
metabolism,  reviews  the  results  of  earlier  investigations.  The  follow- 
ing summary  is  taken  from  Benedict's  discussion.  The  black-faced 
type  signify  the  quotient  N:P205. 


PHOSPHORUS  METABOLISM 


401 


DAILY    PHOSPHORUS     (P205)     EXCRETION    AND    RATIO  BETWEEN 
NITROGEN  AND  PHOSPHORUS  EXCRETION  IN  URINE  OF 
FASTING  SUBJECTS— Grams 


Succi 

Breit- 
haupt 

Subject 
I 

Flora 
Tosca 

Day  of 

At 

At 

At 

Cetti 

J 

Sohn 

Keller 

fast 

Florence 

Naples 

Vienna 

4.73 

4.89 

4.84 

7.40 

6.21 

4.53 

5.24 

4.41 

1.90 

2.76 

2.69 

1.682 

3.381 

1.58 

2.670 

1.86 

7.87 

4.90 

5.70 

5.22 

6.42 

5.20 

5.37 

3.97 

5.65 

3.55 

i 

1.930 

1.78 

2.98 

2.597 

1.56 

1.091 

2.303 

2.22 

1.550 

1.90 

5.91 

4.64 

4.07 

4.30 

5.25 

6.51 

5.45 

3.81 

4.58 

3.24 

2 

2.051 

1.82 

2.75 

2.925 

1.89 

1.491 

2.268 

2.65 

1.830 

2.44 

7.30 

4.64 

4.19 

3.99 

5.25 

6.03 

6.17 

4.41 

4.04 

4.54 

3 

2.090 

1.95 

2.52 

3.289 

2.53 

1.571 

2.270 

2.65 

2.654 

2.53 

6.64 

5.83 

4.25 

4.17 

5.42 

5.54 

3.20 

4 

2.120 

1.46 

2.54 

2.974 

2.36 

5.052 

2.934 

5 

5.90 

3.74 

4.46 

3.73 

5.00 

4.50 

2.394 

2.64 

2.51 

2.871 

2.19 

1.749 

5.18 

3.49 

4.85 

3.79 

4.32 

4.43 

7.23 

6 

2.150 

2.47 

2.27 

2.667 

2.29 

2.434 

1.069 

7 

5.53 

3.28 

4.13 

4.09 

4.60 

8.57 

1.865 

2.32 

2.13 

2.663 

3.150 

0.713 

8 

5.85 

4.18 

4.22 

5.17 

4.86 

4.64 

1.601 

1.48 

2.31 

1.722 

4.442 

1.658 

9 

6.29 

4.63 

4.19 

5.24 

4.32 

1.360 

1.49 

2.40 

2.065 

1.702 

10 

5.96 

4.37 

4.24 

10.00 

4.65 

1.246 

1.23 

1.68 

0.948 

1.461 

Av.  ratio 
N<P20e 

6.24 

4.37 

4.43 

4.97 

5.28 

6.29 

5.20 

4.06 

5.14 

3.94 

The  following  table  sets  forth  results  from  Benedict's  investi- 
gation. 


RATIOS  OF  NITROGEN  TO  PHOSPHORUS   (P205)  AND  AMOUNTS 
PHOSPHORUS  EXCRETED  IN  THE  URINE  DURING  FAST 

Grams 


OF 


Exp. 

Subject  and 

First 

Second 

Third 

Fourth 

Fifth 

Sixth 

Seventh 

Initial 

No. 

duration 

day 

day 

day 

day 

day 

day 

day 

body  wU 
Kg-. 

69 

A.  L.  L.,  Dec.  16-19 

10.78 

0.936 

11.96 
1.192 

14.19 
1.060 

12.44 
1.043 

73.806 

71 

S.  A.  B.,  Jan.  7-10 

7.93 

0.736 

8.33 

1.326 

6.55 

2.000 

5.27 

2.038 

58.199 

73 

S.  A.  B.,  Jan.  28-Feb.  1 

4.39 

2.345 

6.29 

1.904 

5.96 

1.938 

4.81 
2.159 

4.68 
2.134 

5.19 
2.071 

4.87 

2.081 

59.134 

75 

S.  A.  B.,  Mar.  4-10 

8.55 
1.431 

5.52 

2.255 

6.34 

2.055 

4.83 

2.406 

5.23 

2.078 

59.520 

77 

S.  A.  B.,  Apr.  8-11 

4.71 

1.871 

3.90 

2.763  ■ 

4.41 

2.492 

4.35 

2.634 

61.612 

79 

H.  E.  S.,  Oct.  13-14 

7.50 

1.081 

8.62    i 
1.664 

57.170 

80 

C.  R.  Y.,  Oct.  27-28 

6.57 

1.184 

4.11 
2.421 

69.342 

81 

A.  H.  M.,  Nov.  21-22 

6.19 

1.472 

5.37 

2.430  ' 

62.016 

82 

H.  C.  K.,  Nov.  24-25 

8.43 
1.113 

7.18    \ 
2.000 

71.493 

83 

H.  R.  D„  Dec.  5-6 

5.51 

2.407 

4.84 

2.793 

55.637 

85 

N.  M.  P.,  Dec.  9-10 

8.79 
1.257 

9.55 

1.189 

67.625 

89 

D.  W.,  Jan.  10-11 

4.24 
2.354 

4.61 

3.134 

79.063 

6.97 

6.69 

5.82 

4.82 

Average 

1.516 

2.089 

1.909 

2.056 

I 

Body  weight  in  Exp.  69-77  includes  underclothing,  Exp.  80  all  clothing,  Exp.  79,  81  and 
82-89,  no  clothing. 


402 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


From  the  two  foregoing  tables  we  note  that  the  phosphorus 
excretion  is  usually  higher  on  the  second  day  than  on  the  first ;  that 
after  the  second  day  there  is  little  regularity  in  the  variations ;  that 
there  is  a  progressive  decrease  in  the  factor  N  :P205,  at  least  during 
the  first  four  days,  which  may  be  due  to  autolysis  of  bone  tissue, 
and  that  the  daily  elimination  of  phosphorus  (P205)  in  the  urine  of 
the  fasting  man  is  about  2.00  grams. 

Benedict  included  in  his  study  of  fasting  metabolism  balance 
determinations  in  the  period  following  the  fast.  The  following 
nitrogen  and  phosphorus  data  exhibit  marked  initial  retention, 
followed  by  progressive  decrease  to  the  normal. 

AVERAGE  DAILY  NITROGEN  AND  PHOSPHORUS   (P205)   BALANCES 
WITH  HUMAN  SUBJECTS  FOLLOWING  A  FAST— Grams 


Week  No. 

Nitrogen 

Phosphorus  (P2O5) 

Exp.  No. 

Food 

Urine 

Feces 

Balance 

Food 

Urine 

Feces 

Balance 

1 
1 
1 
1 

1 
2 
3 
4 

21.11 
21.80 

17.38 
15.67 

10.60 
10.56 
11.40 
11.86 

2.35 
3.88 
2.79 
2.54 

+8.17 
+7.36 
+3.19 
+1.27 

10.712 
8.699 
5.899 
5.588 

1.804 
2.673 
3.097 
2.795 

2.015 

2.787 
1.983 
1.865 

+6.893 
+3.238 
+0.819 
+0.928 

2 
2 

1 

2 

19.61 
19.41 

14.13 

12.48 

2.46 
2.36 

+3.02 

+4.57 

5.889 
5.907* 

2.256 
2.913* 

2.235 

1.985* 

+1.398 
+1.009 

*  For  six  days. 

Phosphorus  determinations  on  urine  of  Exp.  I  were  made  by  the 
uranium  acetate  method ;  those  of  Exp.  2  were  made  by  fusion. 

Halpern  (1908)  obtained  data  as  follows  on  the  urinary  excre- 
tion of  a  man  who  was  about  to  die  with  cancer.  The  urine  for 
two  days,  during  which  no  nutriment  other  than  30  gm.  grape  sugar 
in  600  c.c.  of  water  was  taken,  contained  the  following  constituents 
per  day:  Total  N,  2.058  gm.  (after  deducting  albumin,  2.0097  gm.) ; 
ammonia  N,  0.0714  gm.,  or  3.47  percent  of  the  total  N;  purin  N, 
0.059  gm. ;  S03,  0.344  gm.,  and  P205,  0.319  gm.  The  ratio  of  N  to 
P205,  therefore,  was  as  6.3:1. 

Summary.  The  phosphorus  excretion  of  an  adult  human 
being  during  fast  is  about  2.00  gm.  P205;  the  excretion  varies  in 
general  with  the  loss  in  weight.  There  is  in  fast  a  progressive 
decrease  in  acid  and  alkaline  phosphates  in  the  urine;  the  earthy 
phosphates  show  first  an  increase,  and  subsequently  a  return  to  the 
earlier  figures.  There  is,  at  least  during  the  first  four  days  of  fast, 
a  progressive  decrease  in  the  proportion  of  nitrogen  to  phosphorus 
in  the  urine.      The  phosphorus  excretion  immediately  subsequent  to 


PHOSPHORUS  METABOLISM 


403 


fast  may  be  much  less  than  during  fast.  The  retention  of  phos- 
phorus after  the  cessation  of  fast  is  very  marked  but  gradually 
returns  to  the  normal. 

For  a  discussion  of  other  phases  of  phosphorus  metabolism 
during  fast  see  P.  Met.  During  Fast. 

NORMAL  PHOSPHORUS  REQUIREMENTS  OF  ADULT  HUMAN  BEINGS 

Siewert  (1868)  publishes  results  of  nitrogen  balance  experi- 
ments with  human  subjects,  balance  data  on  inorganic  elements  also 
being  included  in  some  cases.  A  part  of  the  results  are  set  forth 
below. 

AVERAGE  DAILY  NITROGEN,  CALCIUM  AND  PHOSPHORUS  BALANCES 
WITH  A  HUMAN  SUBJECT  ON  A  NORMAL  MIXED  DIET— Grams 


Experiment 

No. 

N 

Food 

Urine 

Feces 

Balance 

CaO 

Food 

Urine 

Feces 

Balance 

P05 

Food 

Urine 

Feces 

Balance 

Body  weight        , 

Initial 
Final 

2 
Feb.  10-19 

19.051 

15.844 

2.788 

+0.419 

1.093 

0.209 

0.963 

—0.079 

4.932 

3.265 

1.740 

-0.073 

56000 
55450 

3 
Mar.  2-11 

19.360 

15.682 

2.563 

+1.115 

1.026 

0.183 

0.880 

-0.037 

5.441 

3.118 

2.381 

-0.058 

55600 
56750 

The  negative  phosphorus  balances  must  have  been  due  to  a 
well-established  habit  of  excessive  phosphorus  intake  and  outgo, 
because  the  intake,  in  amounts  as  above  stated,  is  usually  ample  for 
maintenance  purposes. 

Piatt  (1897)  submits  the  following  data  for  the  average  daily 
urinary  phosphorus  of  adult  Americans.  The  figures,  of  course, 
represent  food  habit  more- accurately  than  food  requirement. 

PHOSPHORUS  IN  NORMAL  HUMAN  URINE— Grams  in  24  Hours 


Normal  urine 


Man 


Woman 


Total  phosphorus  (P2O5) . 
Glycerophosphoric  acid. . . 


2.0-3.5 
0.01—0.02 


3.0 
0.015 


2.5 


See   also   Krabbe    (1857). 

Siven  (1901)  reported  results  of  metabolism  experiments  on  a 
man  who  was  31  years  old,  and  who  weighed  normally  about  65.2 
kg.,  with  rations  of  normal  foods  so  chosen  as  to  vary  the  protein 
content  of  the  diet.  A  portion  of  the  numerical  data  are  given 
below. 


404 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


DAILY  PHOSPHORUS  BALANCES  OF  A  MATURE  MAN  ON  RATIONS 
VARYING  IN  PHOSPHORUS  CONTENT— Grams 


Date 

Food 

Urine 

Feces 

Bal- 
ance 

Urine 

and 

Feces 

Diet 

Nov.  14 
*'    21 

0.526 
0.526 

0.458 
0.356 

0.283 
0.331 

0.741 

0.687 

-0.215 
—0.161 

Potatoes,  apples,  butter,  sugar,  beer 

"    22 
"    23 
"    24 
"    25 

0.644 
0.644 
0.644 
0.644 

0.389 
0.381 
0.437 
0.523 

0.364 
0.364 
0.364 
0.364 

0.753 
0.745 
0.801 
0.887 

-0.109 
-0.101 
-0.157 
-0.243 

"             "            "            "        «'        eggs 

"    26 

"    27 

"    29 

Dec.    1 

•■      2 

1.131 
1.653 
1.653 
1.629 
1.558 

0.654 
0.887 
0.978 
0.802 
0.964 

0.460 
0.460 
0.460 
0.460 
0.460 

1.114 

1.347 
1.438 
1.262 
1.424 

+0.017 
+0.306 
+0.215 
+0.367 
+0.134 

::     ::     ::'     ::    ::     :: 

"      3 

-  i 

"      6 
.       ? 

"      8 

3.183 
3.124 
3.124 
3.099 
3.194 
3.147 

1.108 
1.410 
1.441 
1.410 
1.445 
1.441 

1.595 
1.595 
1.595 
1.595 
1.595 
1.595 

2.703 
3.005 
3.036 
3.005 
3.040 
3.036 

+0.480 
+0.119 
+0.088 
+0.094 
+0.154 
+0.111 

Bread,  milk,  eggs,  cheese,  "butter,  apples,  beer 
it          ii        <i            •«            ii            it        •« 

If                    11                II                       II                       II                       14                •« 

From  these  data  the  author  concludes  that  the  minimum 
daily  requirement  of  phosphorus  of  the  adult  human  being  is  about 
0.7-0.8  gm.  From  the  data  of  Nov.  22-Dec.  2  we  compute  that  with 
this  subject  the  requirement  was  more  than  0.8  and  less  than  1.3 
gm.  P. 

Ehrstrom  (1903b)  succeeded  in  maintaining  phosphorus  bal- 
ance during  13  days  in  a  woman  40  years  old  solely  by  the  use  of 
rectal  injections  of  milk  and  a  casein-containing  preparation, 
"proton,"  the  phosphorus  intake  per  day  being  0.61  gm.  during  8 
days,  and  0.91  gm.  during  the  last  5  days.  The  phosphorus  absorbed 
during  the  first  8  days  was  0.42  gm.  per  day,  and  during  the  last  5 
days  0.74  gm.  per  day. 

In  another  case,  a  man  42  years  old,  the  injection  of  1.22  gm. 
P,  and  absorption  of  0.51  gm.,  did  not  maintain  equilibrium ;  loss  per 
day  0.530  gm.  P. 

A  third  case,  a  woman  31  years  old,  was  not  maintained  in 
phosphorus  equilibrium  by  a  rectal  intake  of  1.22  gm.  P  per  day, 
and  an  absorption  of  0.71  gm. ;  loss  per  day  0.19  gm.  P. 

The  result  with  the  first  mentioned  case  suggests  the  high 
efficiency  of  casein  phosphorus. 

Maurel  (1901,  1904)  in  experiments  on  himself,  with  normal 
diet,  concluded  that  the  phosphorus  requirement  of  the  human 
being  is  0.04-0.05  gm.  P205  per  kg.  of  body  weight.  In  this  connec- 
tion he  considers  also  the  content  of  P2O0  in  human  milk,  which  is 


PHOSPHORUS  METABOLISM  405 

stated  to  be  0.05  gm.  P205  per  100  gm.  of  milk,  which  is  about  the 
amount  consumed  by  infants  per  kilogram  of  body  weight  per  day. 
Since  Maurel's  experiments  had  to  do  with  urinary  excretion  alone 
they  hardly  furnish  a  basis  for  a  close  estimate  of  the  phosphorus 
requirement. 

In  balance  experiments  on  a  diet  of  crackers,  butter  and  milk, 
with  a  man  weighing  60  kg.,  Sherman  (1902)  found,  in  4-  and  5-day 
periods,  that  in  two  experiments  there  was  a  negative  balance  on  an 
intake  of  1.58  gm.  phosphorus;  again  a  negative  balance  on  an 
intake  of  1.4  gm.  phosphorus,  and  a  positive  balance  on  an  intake  of 
1.6  gm.  phosphorus.  These  amounts  appear  to  be  close  to  the 
phosphorus  requirement  under  normal  conditions. 

From  the  work  of  Ehrstrom  (1903a)  (see  p.  322)  it  would 
appear  that  under  the  conditions  of  the  second  period  of  his  experi- 
ment phosphorus  equilibrium  could  have  been  maintained  on  less 
than  1.348  gm.  P. 

Renvall  (1904),  experimenting  on  himself,  at  an  age  of  22  years 
and  weight  of  71.1  kg.,  found  in  a  series  of  balance  experiments  with 
a  normal  mixed  diet,  that  in  a  5-day  period  the  average  daily  phos- 
phorus loss  was  0.030  gm.  on  an  intake  of  2.103  gm.  of  P205,  the 
most  that  was  fed  during  any  period  in  the  series.  The  requirement, 
therefore,  seems  to  have  been  more  than  2.133  gm.  P205. 

Gilbert  and  Posternak  (1903)  failed  to  maintain  P205  equi- 
librium on  an  intake  of  2.43  gm.  daily,  but  with  an  addition  of 
lecithin,  bringing  the  intake  up  to  2.745  gm.,  as  an  average  of  a 
3-day  period,  there  was  a  retention  of  0.031  gm.  P205  per  day;  thus 
under  these  conditions  the  phosphorus  requirement  was  apparently 
less  than  2.714  gm.  P205. 

In  the  course  of  their  investigations  of  the  influence  of  food 
preservatives  and  artificial  colors  on  digestion  and  health,  Wiley, 
and  associates  (1904,  1906,  1907,  1908a,  b)  accumulated  a  large 
amount  of  balance  data  which  are  not  without  value  as  indications  of 
the  phosphorus  requirements  of  human  beings.  The  experiments 
were  all  on  healthy  young  men,  and  for  the  purpose  of  this  discus- 
sion we  submit  average  daily  balances  of  the  groups  used  in  the 
several  investigations,  the  data  covering  the  fore-periods  only,  dur- 
ing which  no  preservatives  were  fed. 

One  circumstance  requires  mention  as  bearing  on  the  accuracy 
of  these  balances.  The  feces  were  not  marked  at  the  beginning 
and  end  of  the  experimental  periods.  The  large  number  of  subjects 


406 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


and  periods,  however,  tends  to  reduce  the  error,  though  the  periods, 
being  5-10  days  in  duration,  were  too  short  to  be  really  satisfactory 
for  our  purpose.      The  food  was  normal  and  varied  in  character. 

AVERAGE  DAILY  PHOSPHORUS  BALANCES  WITH  HEALTHY  YOUNG 

Men — Grams  P205 


Number   of 
subjects 

Number  of 
periods 

Body 

weight 

Kg. 

Food 

Urine 

Feces 

Balance 

Publication 

12 
9 
12 
11 
10 

30 

9 

12 

11 

10 

66.14 
62.71 
63.69 
63.27 
63.30 

4.358 
3.816 
4.312 
3.782 
3.956 

2.648 
2.364 
2.455 
2.244 
2.375 

1.571 
1.247 
1.352 
1.047 
1.262 

+0.139 
+0.205 
+0.508 
+0.491 
+0.319 

Bu.  Chem.  Bui.  84,  Part   I 
"     '•      "      II 
"     "      "     III 
....       «•     IV 

V 

These  data  show  that  a  free  choice  of  normal  foods  provides  a 
considerable  excess  of  phosphorus,  and  that  these  young  men  of 
somewhat  under  average  weight  naturally  took  in  the  food  3.78-4.36 
gm.  of  P205. 

VonWendt  (1905)  finds  that  if  the  human  body  receives  a 
sufficient  amount  of  oxidizable  nitrogen-free  material,  the  outgo 
of  nitrogen,  sulphur  and  salts  may  be  considerably  decreased, 
through  the  establishment  of  an  internal  circulation  by  means  of 
which  these  substances  are  retained  in  the  body  and  are  available 
for  repeated  use.  VonWendt  states  that  the  body  maintenance 
requirement  of  phosphorus  (P)  is  about  0.010  gm.,  of  calcium  0.008 
gm.,  and  of  magnesium  not  more  than  0.001  gm.  per  kilogram  of 
body  weight. 

Experiments  of  Hawk  (1905),  with  two  normal  mature  men 
weighing  61.4  and  62.8  kg.,  on  a  diet  of  crackers,  butter  and  milk 
showed  that  with  an  intake  of  4.96  gm.  P205  daily  there  resulted  a 
storage  of  0.145,  0.279  and  0.207  gm.  P205  in  2-day  periods.  These 
individuals  were  living  on  a  plane  of  phosphorus  metabolism  which 
did  not  require  economy  by  the  body. 

With  a  30-year-old  man  Gilbert  and  Posternak  (1905)  failed  to 
maintain  phosphorus  equilibrium  on  an  intake  of  3.19  gm.  P205 
daily,  but  got  a  storage  of  0.121  gm.  P205  on  an  intake  of  3.43 
gm.  P2Og  daily,  the  requirement  thus  appearing  to  be  less  than  3.31 
gm.  P,O0. 

In  experiments  by  Gumpert  (1905)  an  adult  man  lost  0.018  gm. 
P205  per  day  on  an  intake  of  1.885  gm.  in  a  5-day  period,  the  phos- 
phorus requirement,  therefore,  appearing  to  be  greater  than  1.903 
gm.  P205.  The  diet  was  of  normal  mixed  foods  plus  casein.  In 
other  periods,  containing  meat  instead  of  casein,  the  phosphorus 
requirement  seems  to  have  been  much  greater. 


PHOSPHORUS  METABOLISM 


407 


E.  Koch  (1906)  studied  phosphorus  metabolism  in  a  man  weigh- 
ing 63.9  kg.  On  a  normal  mixed  ration,  in  a  four-day  period,  with 
a  phosphorus'  intake  of  4.43  gm.  P205  daily,  the  storage  was  0.34  gm. 
daily,  the  intake,  therefore,  being  too  much  in  excess  of  the  mini- 
mum to  afford  critical  evidence  as  to  actual  requirement. 

Robert  and  Parisot  (1906)  quote  from  the  tables  of  Roche,  et  at. 
the  following  statement  of  the  normal  phosphorus  content  of  the 
urine  of  people  from  middle  life  to  old  age.  These  figures  reflect 
especially  the  diminished  food  consumption  accompanying  decreased 
activity. 

PHOSPHORUS  OF  NORMAL  HUMAN  URINE 


Age 

Grams  P2O5  for  24  hours 

54-59 
60-64 
65-69 
80-85 

1.85 
1.46 
1.35 
1.0-1.10 

Hamatainen  and  Helme  (1907)  conducted  a  series  of  one-day 
balance  observations  with  rations  of  very  low  phosphorus  content. 
The  daily  phosphorus  balances  are  as  follows : 

PHOSPHORUS   METABOLISM   ON   A   LOW-PHOSPHORUS   DIET   WITH 

SUPPLEMENTS  ADDED  AT  INTERVALS 

Daily  Balances  in  Grams  P 


Day 

Food 

Urine 

Feces 

Balance 

Remarks 

1 

0.377 

0.683 

0.244 

-0.550 

2 

0.324 

0.666 

-0.586 

3 

0.524 

0.639 

-0.359 

4 

0.586 

0.630 

-0.288 

5 

0.517 

0.427 

-0.154 

6 

0.536 

0.407 

-0.115 

7 

0.705 

0.350 

+0.153 

800  gm.  egg  white=0.114  gm. 

P 

8 

0.604 

0.418 

0.202 

-0.046 

9 

0.573 

0.436 

-0.065 

10 

0.567 

0.367 

-0.002 

11 

0.551 

0.512 

-0.163 

12 

0.559 

0.366 

-0.009 

13 

0.552 

0.439 

-0.089 

14 

0.565 

0.443 

1  * 

-0.080 

15 

0.554 

0.401 

4  * 

—0.049 

16 

1.040 

0.663 

0.255 

+0.122 

57  gm.  proton=0.479  gm.  P 

17 

0.559 

0.368 

<■ 

-0.064 

18 

0.552 

0.390 

-0.093 

19 

0.557 

0.393 

-0.091 

20 

1.186 

0.739 

0.233 

+0.214 

320  gm.  roast  veal=0.627  gm 

P 

21 

0.557 

0.518 

.. 

-0.194 

22 

0.555 

0.473 

' ' 

-0.151 

23 

0.535 

0.355 

-0.053 

408  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

On  the  strength  of  the  records  of  the  8th,  10th  and  12th  days 
the  authors  state  that  the  body  is  able  to  maintain  a  phosphorus 
balance  on  0.55-0.65  gm.  phosphorus  per  day.  We  would  suggest, 
however,  that  on  12  other  days,  with  an  intake  of  0.55-0.60  gm., 
there  was  decided  loss  of  phosphorus.  These  figures  show  such  a 
daily  variation  in  phosphorus  outgo  that  we  may  be  warned  against 
drawing  conclusions  from  single  day's  balances. 

In  experiments  by  Oeri  (1909),  a  man  25  years  old,  weighing 
93  kg.,  and  engaged  in  laboratory  work,  failed  to  maintain  phospho- 
rus equilibrium  on  6.54  gm.  P205  intake  daily  in  a  normal  mixed  diet. 
The  addition  of  2  gm.  P205  as  disodium  phosphate  resulted  in 
marked  storage.  On  approximately  the  same  kind  of  diet  a  woman 
35  years  old,  and  weighing  55  kg.,  stored  something  like  half  of  a 
gram  of  P2Og  daily  on  an  intake  of  5.45  gm.  P205.  Neither  of  these 
subjects  was  using  phosphorus  economically. 

Hoifstrom  (1910),  in  studying  metabolism  in  pregnancy,  deter- 
mined that  during  24  weeks,  the  17th-40th  of  pregnancy,  with  an 
average  daily  intake  of  1.952  gm.  phosphorus,  the  average  daily 
storage  was  0.331  gm. 

Sherman,  Mettler  and  Sinclair  (1910),  in  a  series  of  6  three-day 
balance  experiments  on  a  healthy  man  studied  calcium,  magnesium 
and  phosphorus  metabolism.  A  portion  of  the  results  are  in  the 
following  table  (p.  409). 

Nitrogen  on  feces  was  estimated  from  previous  work. 
The  two  rations  which  came  nearest  to  maintaining  phosphorus 
equilibrium  were  Nos.  1  and  6  where,  with  incomes  of  1.546  gm. 
and  1.509  gm.,  respectively,  the  loss  was  0.054  gm.  and  0.015  gm.  of 
phosphorus,  the  requirement,  therefore,  apparently  being  more  than 
the  intake  plus  the  loss,  or  1.6  gm.  and  1.524  gm. 

The  authors  conclude :  "From  the  results  here  obtained,  as  well 
as  the  average  results  of  experiments  by  other  observers,  it  would 
appear  that  a  healthy  man,  accustomed  to  full  diet  of  the  ordinary 
mixture  of  animal  and  vegetable  food  materials,  requires  for  the 
maintenance  of  his  ordinary  store  of  phosphorus  compounds  about 
1.5  gm.  of  phosphorus,  or  nearly  3.5  gm.  of  P205  per  day,  though 
under  special  conditions  or  with  a  specially  selected  dietary,  equi- 
librium may  be  maintained  on  much  less,  (0.9  gm.  phosphorus  or 
about  2.0  gm.  PA)-" 


PHOSPHORUS  METABOLISM 


409 


AVERAGE  DAILY  BALANCES  OF  NITROGEN,  CALCIUM,  MAGNESIUM 
AND  PHOSPHORUS  WITH  AN  ADULT  MAN  ON  A  SIMPLE  DIET 

Grams 


Experi- 

Initial and 

Fuel  value 

N 

Ca 

Mgr 

P 

ment 

final  body 

weight 

Food 

Food 

Food 

Food 

Urine 

Urine 

Urine 

Urine 

Diet 

Feces 

Feces 

Feces 

Feces 

No. 

Lbs. 

Cal. 

Balance 

Balance 

Balance 

Balance 

10.10 

2.651 

0.286 

1.546 

150  gm.  crackers,  1500 

13.09 

0.210 

0.190 

1.030 

firm,  milk 

0.46 

1.880 

0.170 

0.570 

1 

143-137.5 

1690 

—3.45 

+0.561 

-0.074 

-0.054 

10.69 

0.139 

0.113 

0.384 

400  gm.  crackers,  250 

13.21 

0.097 

0.130 

0.753 

g-m.       coagulated 

0.75 

0.480 

0.080 

0.223 

white  of  egg: 

2 

137.5-137.5 

1833 

-3.27 

-0.438 

—0.097 

-0.592 

7.02 

0.126 

0.079 

0.401 

450  gin.  crackers 

10.29 

0.054 

0.093 

0.695 

0.70 

1.230 

0.126 

0.452 

3 

138-136 

1930 

-3.97 

-1.158 

-0.140 

-0.746 

9.41 

0.948 

0.160 

0.836 

450  grm.  crackers,  450 

9.95 

0.108 

0.132 

0.659 

erm.  milk,  75    gm. 

0.77 

0.716 

0.053 

0.226 

butter 

4 

136-139 

2774 

-0.91 

+0.124 

-0.025 

-0.049 

7.00 

0.866 

0.132 

0.702 

300  grm.  crackers,  450 

8.54 

0.188 

0.138 

0.790 

grm.  milk,  75    grm. 

0.54 

0.582 

0.046 

0.160 

butter 

5 

139-137.5 

2140 

-2.08 

+0.096 

-0.052 

—0.248 

11.64 

2.237 

0.263 

■ 
1.509 

300  g-m.  crackers,  1350 

10.59 

0.307 

0.179 

0.992 

grm.  milk 

0.68 

1.807 

0.141 

0.502 

6 

137.5-137 

2170 

+0.37 

+0.123 

-0.057 

—0.015 

Sherman  et  al.  (1910)  compute  the  following  estimates  of  the 
ash  constituents  of  typical  American  dietaries. 

ESTIMATED  AVERAGE  DAILY  QUANTITIES  PER  MAN  OF  CERTAIN 
CONSTITUENTS  IN  TYPICAL  AMERICAN  DIETARIES— Grams 


Persons  studied 


Maine  lumbermen 

School  superintendent's  family,  Chicago 

Student's  club,  Univ.  of  Tennessee 

Decorator's  family,  Pittsburgh , 

Farmer's  family,  Connecticut , 

Teacher's  family,  Indiana 

Teacher's  family,  New  York  City 

Mechanic's  family,  Tennessee 

Farmer's  and  mechanic's  family,  Tennessee 

Glassblower's  family,  Pittsburgh 

Lawyer's  family,  Pittsburgh 

Women  students'  club,  Ohio 

Laborer's  family,  New  York  City 

Laborer's  family,  Pittsburgh 

Negro  farmer's  family,  Alabama 

Laborer's  family,  Pittsburgh 

Laborer's  family,  New  York  City 

Farm  students'  club,  Tennessee 

Sewing  woman's  family,  New  York  City.. . . . 
Very  poor  negro  family,  Alabama 


Protein 

Fe 

P2O5 

CaO 

MgO 

Cal. 

6780 

179 

0.035 

5.88 

1.27 

1.21 

3260 

123 

0.021 

3.97 

1.09 

0.55 

3595 

123 

0.019 

4.05 

1.22 

0.63 

3305 

112 

0.019 

3.44 

0.90 

0.48 

3545 

108 

0.021 

3.53 

1.15 

0.55 

2780 

106 

0.016 

3.64 

1.42 

0.44 

3180 

102 

0.016 

3.92 

1.69 

0.54 

4060 

97 

0.017 

3.58 

0.90 

0.72 

2820 

95 

0.019 

3.56 

0.83 

0.59 

3085 

94 

0.016 

2.73 

0.49 

0.36 

3280 

91 

0.015 

2.82 

0.83 

0.40 

3330 

85 

0.015 

2.88 

0.97 

0.67 

2335 

84 

0.014 

2.41 

0.47 

0.30 

2525 

83 

0.013 

2.40 

0.50 

0.34 

4955 

80 

0.012 

3.25 

0.21 

0.74 

2440 

77 

0.012 

1.52 

0.40 

0.19 

2430 

71 

0.012 

2.27 

0.50 

0.29 

3560 

66 

0.011 

2.08 

0.46 

0.34 

1500 

64 

0.009 

1.84 

0.68 

0.23 

2240 

44 

0.007 

2.05 

0.08 

0.52 

410 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


The  authors  comment  on  these  figures  in  part  as  follows :  "Many 
of  the  dietary  studies  show  so  much  less  than  3.5  gm.  of  P205  per 
man  per  day  as  to  raise  a  question  whether  these  people  may  not 
have  been  undernourished  in  this  respect,  even  though  they  may 
have  had  ample  proteins,  fats  and  carbohydrates." 

Holsti  (1910),  experimenting  on  himself  at  a  weight  of  58  kg., 
and  age  of  21  years,  retained  0.007  gm.  P  under  normal  conditions 
on  an  intake  of  2.57  gm.  daily  during  a  3-day  period.  With  a  sub- 
normal nitrogen  intake,  which  gave  a  minus  nitrogen  balance,  he 
retained  0.185  gm.  P  on  an  intake  of  1.922  gm.  daily  in  a  6-day 
period,  the  requirement  in  the  latter  case  appearing  to  be  less  than 
1.737  gm. 

Aron  and  Hocson  (1910)  studied  phosphorus  metabolism  in 
connection  with  beriberi.  From  experiments  on  two  normal  indi- 
viduals it  was  determined  that  a  diet  consisting  of  fish,  bread,  rice, 
sugar,  etc.,  furnishing  37  calories,  0.2  gm.  nitrogen,  and  0.032  gm. 
P205  per  kilo  of  body  weight,  is  sufficient  to  keep  a  man  in  N  and 
P205  equilibrium. 

VonWendt  (1910),  in  a  series  of  1-day  balance  experiments  on 
himself,  at  different  altitudes  in  the  Alps,  seems,  on  a  number  of 
days,  to  have  retained  phosphorus  on  an  intake  of  0.861  gm.  P,  but 
on  a  greater  number  of  days  there  were  losses  of  phosphorus  greater 
in  magnitude  than  the  previously  mentioned  storage.  We  must 
conclude  therefore,  that  under  his  conditions  0.861  gm.  P  was  an 
insufficient  amount.  The  calcium  balances  were  negative,  and  the 
nitrogen  balances  positive. 

R.  Tigerstedt  (1911)  studied  the  composition  of  the  freely- 
chosen  food  consumed  by  64  persons  ranging  in  age  from  2  years  to 
maturity.      Some  of  his  results  are  set  forth  in  the  following  table. 

AVERAGE  DAILY  CALCIUM,  MAGNESIUM  AND  PHOSPHORUS  OF  THE 
FREELY  CHOSEN  DIETS  OF  HUMAN  BEINGS  OF  VARIOUS  AGES 

Grams 


Children  2-3  years. 
Children  4-7  years. 
Children  8-11  years 
Girls  12-16  years. . . 
Boys  12-16  years. . . 

Adult  women 

Adult  men 


P2O5 


1.18 
1.93 
2.55 
3.09 
3.09 
2.77 
4.33 


1.58 
1.84 
2.37 
2.21 
2.01 
2.29 
3.79 


MsrO 


0.24 
0.39 
0.59 
0.89 
0.71 
0.66 
1.09 


Tigerstedt  states  his  opinion  that  the  P2Os  intake,  in  order  to 
maintain  the  phosphorus  balance,  should  be  from  2.0  to  3.5  gm.  per 
day.  From  his  investigations  it  appears  that  the  adult  man's  diet 
contains  more  than  this  amount  (mean  4.33  gm.),  while  adult  women 
at  moderate  labor  consume  3.57  gm.  daily.      The  phosphorus  values 


PHOSPHORUS  METABOLISM 


411 


varied  within  wide  limits  (P  for  men  2.80-6.01  gm.,  women,  1.69-4.26 
gm.). 

In  connection  with  their  study  of  rice  as  the  cause  of  beriberi, 
Aron  and  Hocson  (1911a,  1911b)  report  the  following  experiments 
with  healthy  men. 

PHOSPHORUS  BALANCES  ON  HEALTHY  MEN 


Length 

No.  of 

of 

period 

period 

in  days 

3 

i 

4 

4 

5 

4 

6 

4 

7 

4 

8 

4 

9 

6 

10 

4 

11 

4 

12 

3 

13 

3 

14 

4 

15 

4 

Foodstuffs 


Ricel,  sugar,  bacon,  coffee,  onions,  fat 

Rice2,  bread,  sugar,  bacon,  coffee,  onions,fat 

Rice,  bread,  sugar,  bacon,  coffee 

Rice,  bread,  sugar,  bacon,  coffee,  phytin. 
Rice,  bread,  sugar,  bacon,  coffee,  egg  white 

Rice,  fish,  sugar,  coffee,  bananas 

Rice,  bread,  fish,  sugar,  coffee 

Rice,  bread,  fish,  sugar,  coffee , 

Rice,  bread, fish,  sugar,  coffee,  wheat  flour 

Rice,  bread,  fish,  sugar,  coffee 

Rice,  bread,  fish,  sugar,  coffee 

Rice,  bread,  fish,  sugar,  coffee 

Rice,  bread,  fish,  sugar,  coffee 


Calor- 
ies per 

kg. 
body 

wt. 


P2O5 
intake 
per  kg. 

body 
wt. 


.066 
.029 
.023 
.077 
.029 

.'033 
.032 
.104 
.033 
.033 
.032 
.031 


P2O5 
of  urine, 
percent 

of 
intake 


32.0 
73.2 
100.0 
31.7 
83.5 

62!  5 
72.6 
25.3 
57.6 
57.6 
65.5 
74.2 


P2O5 
of  feces, 
percent 

of 
intake 


63.1 

58.4 
48.7 
37.7 
32.4 

38. 6 
33.7 
63.4 
42.9 
41.2 
40.0 


Body 
wt. 


Kg. 


49.0 
48.6 
64.0 
64.0 
64.0 
54.0 
43.5 
52.5 
52.5 
54.0 
54.0 
45.9 
53.0 


P2OS 
balance 


Grams 


+0.16 
-0.45 
-0.97 
+1.50 
-0.30 

-o.'oi 

+0.013 

+0.62 

-0.22 

-0.10 

-0.08 


(1)  Red  rice,  that  is,  only  hulled. 

(2)  White  rice,  overmilled. 

(3)  — 0.09  according  to  our  calculations. 

Authors'  conclusions: 

An  intake  of  1.65  gm.  P205  per  50  kg.  body  weight,  or  less,  does 
not  cover  the  body  requirement.  The  body  loses  phosphorus  on  a 
white  rice  diet,  but  gains  on  the  "red"  rice  (hulled  only). 

Herbst  (1912)  conducted  balance  experiments  on  7  normal  boys 
6-14  years  old.  The  phosphorus  consumption  was  3  gm.  P205  per 
day,  of  which  half  was  recovered  in  the  urine,  and  one-fifth  in  the 
feces;  the  remainder  (0.5-1.0  gm.  per  day)  was  retained.  The 
calcium  and  phosphorus  balances  from  6  of  these  boys,  on  an 
ordinary  mixed  diet,  in  abundance,  are  in  the  following  table  (p. 
412).      The  data  represent  averages  per  day  for  6-day  periods. 

Herbst  (1913)  studied  growth  in  two  boys  aged  about  14  years. 
On  a  normal  mixed  diet,  controlled  by  weights  and  analyses,  the 
boys  were  subjected  to  a  5-months  period  of  observation  on  growth 
and  weight,  in  the  midst  of  which  was  a  12-day  balance  experiment 
during  the  first  6  days  of  which  the  subjects  took  long  walks,  and 
during  the  last  6  of  which  they  took  only  normal  exercise.  On  p.  413 
are  a  part  of  the  balance  data,  which  we  have  calculated  to  average 
figures  per  day. 

During  ordinary  activity  phosphorus  was  stored  in  considerable 
amounts,  the  intake  being  0.073  and  0.067  gm.  P,05  per  kilogram  of 
body  weight.  With  more  liberal  intake  of  calcium  the  phosphorus 
retention  might  have  been  greater. 


412 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  § 


W 
H 


02   g 

<  1 

^  s 

«3 


o 

Q 

iz; 


«! 

P. 
2?  PS 
&  O 


o 

O  CjJ 
O  £ 
«  5 


I* 

P 


d 

tO 

03CM 

t— t— 

TfICO 

inr— 

t-co 

t~Ot 

*+J 

lOTfl 

«o 

CM^I 

CO  CM 

cor- 

00  CO 

r-^-  * — i 

^<H 

Oi— 1 

TTCM 

COtH 

00»— 1 

coo 

■aio 

COO 

ISO 

coo 

CM  CO 

4J 

07 

oo 

oo 

oo 

oo 

oo 

oo 

« 

CO 

CO  CO 

r— r» 

00 1- 

r— m 

Ttiin 

H00 

o 

o 
u 

coco 

OO) 

OlO 

CMH 

cot- 

00  CM 

NN 

HS4CM 

CM  CM 

CO  CM 

t^CM 

coo 

coo 

ISO 

ino 

coo 

TKO 

"3 

fa 

oo 

oo 

oo 

OO 

oo 

oo 

,fi 

o 

O 

^ 

a 

0} 

HO 

OOJ 

f-CM 

Hin 

OlO 

oin 

.3 

.9 

CT>r*- 

oco 

Ht~ 

OiH 

t^co 

CM  CM 

GOO 

COO 

ino 

COCO 

t^o 

COi— 1 

3 

"E 

HO 

HO 

HO 

OO 

o_o 

CMO 

13 

P 

oo 

OO 

OO 

oo 

oo 

oc> 

o 

a) 

r^-tra 

T»<lO 

coco 

Hf~ 

tHCO 

00  CM 

^ 

O^JI 

in  co 

•**t- 

com 

CM0O 

00  05 

rt 

OiT* 

coco 

CDH 

o-** 

r-^si 

CM-HH 

HO 

coo 

OiO 

hO 

oo 

OCO 

a 

HO 

HO 

OO 

HO 

HO 

T-lO 

h 

c 

_o 

CM  CM 

oo 

in  co 

no) 

CO  CM 

o>o 

id 

<M-HH 

TJ<r- 

ooo 

cor- 

CM  CM 

0OCM 

GO"** 

1— (CM 

coco 

oco 

»-HCO 

in  cm 

HO 

I-HO 

coo 

OlO 

t— CO 

TJIO 

HO 

I-HO 

oo 

oo 

OO 

oo 

(0 

D 

U 

0 

O^lO 

C35CM 

HO 

NO 

t-o 

r-H 

rt 

m 

i— ICO 

CDt- 

CMH 

CO  CM 

00 1>- 

in  in 

HOI 

OH 

TTCO 

CO  CM 

CJ5CM 

CM  CM 

d 

CJ 

r~o 

r-o 

coo 

in  co 

mo 

mo 

h 

fa 

oo 

oo 

oo 

oo 

©o 

o'o 

**$ 

O 

CO 

3 

S 

4) 

moo 

om 

(MO 

00  CO 

t^co 

CM  CM 

o 

.2 

OSHJI 

IOO> 

t^t- 

OS  CO 

I— lf- 

CMOS 

,d 

com 

Tti-^fi 

OCO 

OCO 

Iflt* 

o. 

"E 

T*0 

OO 

TXO 

coco 

t-o 

coo 

Cfl 

o 

P 

HO 

MO 

HO 

HO 

HO 

HO 

/a 

PM 

V 

coco 

osr- 

ooo 

Hin 

cm  m 

t^co 

com 

io  co 

coS 

CJilO 

coco 

coco 

COO! 

COOS 

■»HCM 

CM  CO 

cSh 

eon 

ooo 

COH 

CO-— 1 

O"— 1 

a 

coo 

COO 

NO 

coo 

coo 

CMO 

H 

d 

_o 

GOr- 

>— IIO 

r-t~- 

CM-* 

r-oo 

cjiin 

oo 

-M 
a) 

r-i>- 

coh 

COO 

osin 

CO-* 

O— 1 

i 

oo 

HO 

oo 

no 

-*o 

c^o 

HO 

HO 

HO 

HO 

oo 

« 

.  fca 

.  hi 

.  til 

.  ta 

.  hi 

.  bt 

Cfl    M 

%Z 

CO    M 

Cfl    M 

•2  u 

»rt 

ra  aj 

rt  u 

a 

a 

a 

p. 

p. 

p. 

oo 

CM 

CO 

^ 

OJ 

m  tj 

i 

£M 

CO 
CM 

T* 

H 
CM 

CM 

CM 

CM 

o 

CM 

3 

to 

o 

Cfl 

Cfl 

o 

o 

O 

a 

o 

o 

cu 

a 

a 

a 

a 

a 

< 

o 

r- 

CO 

rS 

co 

in 

U9 

Cfl 

Cfl 

w 

at 

CO 

tx 

l>> 

u 

I 

I 

«* 

CO 

00 

r- 

CO 

1-1 

1-1 

H 

PHOSPHORUS  METABOLISM 


413 


AVERAGE  DAILY  NITROGEN,  CALCIUM  AND  PHOSPHORUS  BALANCES 
WITH  TWO  GROWING  BOYS— Grams 


Boy 
No. 

Body  weight 

Initial 
Final 

Kg-. 

N 

Food 

Urine 

Perspiration 

Feces 

Balance 

CaO 

Food 

Urine 

Feces 

Balance 

P2O5 

Food 

Urine 

Feces 

Balance 

Exercise 
taken 

I 

Age 
14  years,  3  mo. 

36.360 
36.335 

36.335 
36.990 

12.357 

10.040 

0.673 

1.177 

+0.467 

14.600 

11.094 

0.217* 

1.800 

+1.650 

0.823 

0.251 

0.291 

+0.281 

0.911 

0.250 

0.507 

+0.154 

2.203 

1.220 

0.436 

+0.547 

2.669 

1.439 

0.719 

+0.511 

Walking-  period 
Normal  activity 

II 

Age 
13  years,  10  mo. 

43.005 
41.740 

41.740 
42.640 

12.469 
12. 162 
0.386 
0.811 
-0.879 

15.507 
12.908 
0.133* 
1.216 
+1.250 

0.829 

0.121 

0.202 

+0.506 

0.953 

0.215 

0.336 

+0.402 

2.224 

1.681 

0.376 

+0.167 

2.826 

1.788 

0.560 

+0.478 

WaUrinjr  period 

i 

Normal  activity 

"Estimated. 

Summary.  From  all  of  these  data  we  conclude  that  the  usual 
phosphorus  requirement  of  an  adult  human  being  of  average  weight 
is  about  1.5-1.75  gm.  P  or  3.4-4.0  gm.  P205 ;  that  under  special  condi- 
tions of  diet  and  previous  feeding  this  amount  may"  be  reduced  to 
about  0.9  gm.  P  or  2.00  gm.  P205. 

Regarding  the  optimum  phosphorus  content  of  the  food  for 
immature  human  beings  beyond  the  age  of  infancy  we  have  only  the 
above  work  of  Herbst. 


414 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


PHOSPHORUS  REQUIREMENTS  OF  INFANTS 

Very  few  balance  experiments  with  normal  infants  are  to  be 
found  in  the  literature  of  the  subject  from  which  it  is  possible  to 
compute  their  phosphorus  requirements.  Those  which  have  come 
to  our  attention  are  here  recorded  in  brief. 

Michel  (1896)  conducted  balance  experiments  with  new-born 
infants  on  a  milk  diet.      A  part  of  the  data  are  as  follows : 

AVERAGE    DAILY    RETENTION    OF    NITROGEN,    CALCIUM    AND 
PHOSPHORUS  WITH  NEW-BORN  INFANTS  ON  A  MILK  DIET 


Age  in  days 

Weight, 

initial  and 

final 

Grams 

Milk 
■received 
per  day 

Grams 

Milk  per 
kg.  body 
weight 

N 

Gain  per 
kg.  body 
weight 

P2O5 

Gain  per 
kg.  body 
weight 

CaO 

Gain  per 

kg.  body 

weight 

5-8 
11-14 
5-9 
7-13 

3730 
3810 

4400 
4520 

2680 
2836 

3500 
3675 

480 
835 
485 
666 

127.3 
187.2 
176.0 
185.0 

0.312 
0.358 
0.402 
0.302 

0.068 
0.063 
0.062 
0.059 

0.055 
0.045 
0.047 
0.054 

3643 

617 

168.9 

0.3435 

0.063 

0.050 

Michel  and  Perret  (1899)  report  that  a  two-and-a-half -months- 
old  infant,  weighing  4725  gm.  at  the  beginning,  gained  19  gm.  per 
day  for  3  days  on  a  daily  intake  of  1.675  gm.  nitrogen,  0.268  gm. 
P205  and  0.377  gm.  CaO,  which  equals  0.35  gm.  nitrogen,  0.056  gm. 
P205  and  0.079  gm.  CaO  per  kg.  body  weight.  The  storage  of  nitro- 
gen was  0.785  gm.,  and  of  P205  0.121  gm.  per  day. 

Blauberg  (1900a,  1900b)  compared  artificial  with  natural  feed- 
ing of  infants.  Balance  data  for  calcium,  magnesium,  and  phospho- 
rus are  as  follows : 

DAILY  CALCIUM,  MAGNESIUM  AND  PHOSPHORUS  BALANCES  WITH 
ARTIFICIALLY  FED  INFANTS— Grams 


Subject 

Diet 

Income 
CaO 

Balance 
CaO 

Income 
MgO 

Balance 
MgO 

Income 
P2O5 

Balance 
P2O5 

3    Normal  (?) 
6%  mo.  old. . . 

Diluted,  sweetened  and 
sterilized  cow's  milk.. 

"Kufeke  mehl"  and 
Sterilized,  undiluted 

.752 

.096 

1.784 

.272 

0.154 

-0.052 

0.924 

0.018 

.102 
.095 
.129 
.044 

0.032 

-0.027 

0.017 

0.018 

.725 

.465 

1.766 

0.203 

0.149 

-0.054 

0.507 

4    Normal 

0.093 

PHOSPHOKUS  METABOLISM  415 

From  these  data  the  author  concludes  that  we  must  consider 
feeding  with  cow's  milk,  even  when  diluted,  to  be  "supernutrition" 
as  regards  the  above  mineral  nutrients,  that  is,  considering  the 
healthy  breast-fed  infant  as  the  standard. 

In  Keller's  experiments  (1900b)  with  infants  on  milk  diet  (see 
table  on  p.  452)  it  was  found  that  on  human  milk  satisfactory- 
gain  in  weight  and  retention  of  nitrogen  and  phosphorus  were 
induced  in  infants  2-5  months  old  (Exp.  2,  6  and  8),  weighing  3300 
to  4350  gm.,  on  an  intake  of  1.512-1.875  gm.  N  and  0.268-0.325  gm. 
P205  daily,  which  is  equivalent  to  361-485  mg.  N  and  67-81  mg.  P205, 
or  29-35  mg.  P,  per  kilogram  of  body  weight. 

In  these  experiments  satisfactory  gain  in  weight  was  not  pro- 
duced with  diluted  cow's  milk.  A  maximum  gain  in  weight  was  pro- 
duced with  whole  cow's  milk,  however,  containing  much  more  than 
the  above-mentioned  amounts  of  nitrogen  and  phosphorus,  with  a 
10-months-old  child. 

From  Netter's  data  (1900)  we  compute  that  6  healthy  infants, 
with  an  average  age  of  8.8  months,  made  the  following  average 
daily  gains  per  kilo  of  body  weight  on  intake  per  kilo  as  stated: 
Intake  of  milk  144  c.c,  of  CaO  0.261  gm.,  and  of  P205  0.335  gm.; 
gain  of  nitrogen  0.204  gm.,  CaO  0.059  gm.,  and  of  P2Og  0.061  gm. 

Rothschild  and  Netter  (1901)  studied  the  effect  of  variation  in 
the  amount  of  milk  taken  by  infants  on  the  thoroughness  of  its 
utilization.  Children  from  4-10  months  old  were  given  milk  in 
amounts  varying  from  125.5-190.2  gm.  per  kg.  of  body  weight.  The 
greatest  gain  of  nitrogen  and  phosphorus  per  day,  per  kg.  of  body 
weight,  occurred  with  a  child  8  months  old,  weighing  7270  gm.,  and 
receiving  177.2  gm.  of  milk  per  kg.  of  body  weight  daily.  On  this 
intake  the  child  gained  0.199  gm.  nitrogen,  0.095  gm.  CaO,  and  0.09 
gm.  P205  per  day,  per  kg.  of  body  weight.  The  composition  of  the 
milk  was  not  given. 

E.  Miiller  (1902)  reported  experiments  with  infants  4  and  6 
months  old  comparing  the  utilization  of  the  constituents  of  raw  and 
sterilized  milk  by  means  of  2  complete  balance  experiments,  each  of 
7  days'  duration,  excreta  being  collected  for  the  last  4  days  of  each 
period.  The  nitrogen  and  fat  of  the  sterilized  milk  were  better 
used  than  of  the  raw  milk.  As  to  calcium  and  phosphorus  the  results 
are  inconclusive.     A  portion  of  the  figures  follow. 


416 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


AVERAGE     DAILY     BALANCES     OF     NITROGEN,     CALCIUM     AND 

PHOSPHORUS  PER  KILOGRAM  OF  BODY  WEIGHT  WITH  INFANTS 

ON  RAW  AND  STERILE  MILK— Grams 


Length  of  period 
in  days 

Treatment  of 
milk 

N 

Food 

Urine 

Feces 

Balance 

P2O5 

Food 

Urine 

Feces 

Balance 

CaO 

Food 

Urine 

Feces 

Balance 

4 
4 
4 
4 

Raw 
Sterile 

Raw 
Sterile 

0.592 
0.403 
0.117 

+0.072 

0.593 

0.357 

0.137 

+0.099 

0.539 

0.424 

0.068 

f  0.047 

0.540 

0.419 

0.067 

+0.054 

0.272 
0.141 
0.116 

+0.015 

0.273 

0.092 

0.149 

+0.032 

0.256 

0.156 

0.047 

+0.053 

0.256 

0.151 

0.060 

+0.045 

0.193 

0.002 

0.166 

+0.025 

0.193 
0.002 
0.181 

+0.010 

0.195 

0.003 

0.133 

+0.059 

0.195 

0.003 

0.133 

+0.059 

Bruck  (1908)  has  given  us  two  balance  experiments  with 
artificially-fed  infants.  The  calcium  and  phosphorus  figures  we 
have  calculated  from  the  author's  data. 

AVERAGE  DAILY  CALCIUM  AND  PHOSPHORUS  BALANCES  WITH 
ARTIFICIALLY  FED  INFANTS— Grams 


Exp. 
No. 

Age 

Initial 

body 

weight 

CaO 

intake 

CaO 

retention 

Percent 

CaO 
retention 

P2O5 
intake 

P2O5 
retention 

Percent 

P2O5 
retention 

Duration 

of 

period 

1 
2 

3  mo. 
8^  mo. 

3400 
5600 

0.602 
0.743 

0.298 
0.299 

45.3 
40.3 

0.720 
1.454 

0.168 
0.300 

23.5 
26.3 

4  days 
3  days 

Thus  with  intakes  of  0.212  and  0.260  gm.  P2Os  per  kg.  of  body 
weight  there  was  retention  of  23.5  and  26.3  percents  of  these 
amounts. 

During  these  tests  these  two  infants  gained  in  weight  40  grams 
and  80  grams  respectively;  the  diet  was  milk,  gruel  and  malt. 

L.  F.  Meyer  (1908)  made  a  study  of  mineral  metabolism  in 
infants  as  affected  by  limited  intake  of  food,  by  increase  in  the 
casein  of  the  food,  and  by  increase  of  the  fat  of  the  food.  Two 
infants  were  the  subjects  of  these  experiments.  "A"  was  3  months 
old,  and  "B"  was  10  months  old  at  the  beginning  of  the  study.  Both 
were  in  a  normal  state  of  general  metabolism,  though  "A"  had  a 
slight  umbilical  hernia,  and  "B"  had  exzema,  and  during  the  interim 
between  the  low  diet  and  the  increased  food  periods  suffered  from 
bronchitis.      A  part  of  the  numerical  data  are  as  follows: 


PHOSPHORUS  METABOLISM 


417 


AVERAGE  DAILY  NITROGEN,  CALCIUM  AND  PHOSPHORUS  BALANCES 

WITH  NORMAL  INFANTS  ON  MILK  DIETS  OF  VARYING 

COMPOSITION— Grams 


Subject 
and 
age 

Length  of 

period  in 

days 

Diet 

N 

Intake 
Urine 
Feces 

Balance 

CaO 

Intake 

Urine 

Feces 

Balance 

P2O5 

Intake 

Urine 

Feces 

Balance 

A 

3  months 

old  at 
beginning 

6 
3 

4 
3 

Low  diet:  200  gm.  milk,  800  gm.  oat  gruel, 
20  gm.  sugar=44  cal.  per  kg. 

High  protein:  2211  gm.  of  1:5  milk,  49  gm. 
casein 

High  protein  and  fat:  2927  gm.  of  1:5  milk, 
80  gm.  salt-free  butter,  66  gm.  casein 

High  fat  diet:  2201  gm.  of  1:5  milk,  60  gm. 
salt-free  butter,  66  gm.  casein 

1.056 
0.88496 
0.1740 
-0.00296 

3.5515 

1.799 

0.2556 

+1.4969 

3.6296 

1.9737 

0.202 

+1.4539 

3.5157 

1.7505 

0.2171 

+1.5481 

0.3485 

0.0242 

0.2657 

+0.0586 

0.3336 

0.0051 

0.2911 

+0.0374 

0.2195 

0.0034 

0.0226 

+0.1935 

0.5686 

0.4003 

0.2772 

-0.1089 

0.7851 

0.4327 

0.1668 

+0.1856 

0.5823 

0.3634 

0.1042 

+0.1147  J 

B 

10  months 

old  at 
beginning' 

3 
8 
3 
4 
3 

Control:  1500  gm.  of  2:3  milk,  1:3  oat  gruel, 
£ 0  gm.  sugar=100  cal.  per  kg. 

Low  diet:  300  gm.  milk,  500  gm.  oat  gruel, 
700  c.c.  water,  60  gm.  sugar=50  cal.  per 
kg. 

High  protein:  4500  gm.  of  1:5  milk,  63  gm. 
casein 

High  protein  and  fat:  6000  gm.  of  1:5  milk, 
100  gm.  salt-  free  butter,  84  gm.  casein 

High  fat  diet:  4500  gm.  of  1:5  milk,  75  gm. 
salt-free  butter,  63  gm.  casein 

5.124 

3.3192 

0.4439 

+1.3609 

2.5690 

3.3603 

0.2232 

-0.0145 

4.7732 

2.947 

0.177 

+1.6492 

4.7732 

3.3439 

0.214 

+1.2153 

4.7732 

3.4137 

0.1815 

+1.1780 

1.3944 
Traces 
1.2674 
+0.1270 

0.4809 

0.0030 

0.4280 

+0.0499 

0.5693 

0.6273 
-0.0580 

0.4270 

0.5468 
-0.1198 

0.5693 

0.6862 
-0.1169 

2.0887 

0.9649 

0.6049 

+0.5200 

0.6695 

0.4451 

0.2306 

-0.0062 

1.1047 

0.5101 

0.5027 

+0.0919 

0.8285 

0.5220 

0.4162 

-0.1097 

1.1180 

0.6483 

0.4452 

+0.0245 

Among  the  author's  conclusions  are  the  following : 

While  losing  in  weight,  on  a  deficient  diet,  a  child  loses  mineral 
matter  and  nitrogen;  after  readjustment  to  the  same  diet,  so  that 
there  is  but  slight  loss  in  weight,  the  loss  in  these  constituents  may- 
be reduced  to  zero. 

Birk's  experiments  (1909)  (see  Rachitis),  with  two  normal 
infants,  showed  that  with  1.55  and  1.51  grams  of  phosphorus  in  the 
food  per  day  there  resulted  a  retention  of  21  and  18  percents  of 
these  amounts. 

Tobler  and  Noll  (1910)  published  mineral  balances  with  a 
healthy  infant  two  and  a  half  months  old.  The  following  table 
sets  forth  the  more  important  results. 


418 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


AVERAGE    DAILY    MINERAL    METABOLISM     WITH     A     HEALTHY 
BREAST-FED  INFANT  TWO  AND  ONE  HALF  MONTHS  OLD— Grams 


Intake 

Urine 

Feces 

Balance 

Percent 
retained 

Retention 

per  day  an 

kilo. 

KO 

0.3432 
0.2214 
0.2390 
0.0284 
0.2102 

0.1124 

0.0078 
0.0372 
0.0184 
0.0571 

0.0582 
0.0085 
0.1485 

0.0343 

+0.1726 
+0.2051 
+0.0533 

+0.1188 

50.27 
92.64 
22.30 

56.51 

0.0431 

Na»0 

0.0513 

CaO 

0.0133 

MgO 

P2O5 

0.0297 

The  milk  contained  in  100  gm. :  ash  0.1596,  K20  0.048,  Na20 
0.0309,  CaO  0.0334  and  P205  0.0294  gm.;  length  of  experiment  6 
days;  initial  weight  4000  gm. ;  final  weight  4170  grams.  From 
these  data  we  conclude  that  there  was  marked  retention  of  phospho- 
rus on  an  intake  of  0.051  gm.  P205,  or  0.022  gm.  P  per  kg.  of  body 
weight. 

Schabad  (1910d)  reported  the  following  figures  for  phosphorus 
excretion  by  children,  which  amounts,  in  relation  to  food  require- 
ments, should,  of  course,  be  considered  as  below  the  minimum. 

AVERAGE  DAILY  PHOSPHORUS  EXCRETION  OF  HEALTHY  CHILDREN 
ON  VARIOUS  DIETS 


Total  output  of  P2O5 
per  kilo,  per  day 


Grams 


Percent 


P2O5  in  urine 
per  kilo, 
per  day 

Grams 


Partition  of  P2O5  output 


Urine 
Percent 


Feces 
Percent 


Naturally  nourished  children 

0.0134 
0.0303 
0.0229 

17 
40 
65 

0.0074 
0.0175 
0.0185 

55.6 
57.3 
80.6 

44.4 

42.7 

19.4 

Artificially  fed  children 

j       0.214 
1       

0.119 

57.5 
65.2 

42.5 
34.8 

Older  children  on  bread  and  milk 

0.159 

0.102 

64.4 

35.6 

Summary.  New-born  infants  (Michel)  weighing  on  an  average 
3643  grams,  and  receiving  of  milk  617  gm.  per  day,  or  168.9  gm.  per 
kilogram  of  body  weight,  retain  about  0.063  gm.  P20D,  or  0.0275  gm. 
P,  per  day ;  the  infant  receiving  cow's  milk  may  retain  considerably 
more  calcium  and  phosphorus  than  the  breast-fed  baby  (Blauberg) ; 
on  human  milk  infants  2-5  months  old  made  satisfactory  retention 
of  phosphorus  on  an  intake  of  29-35  mg.  per  kg.  of  body  weight  per 
day  (Keller) ;  infants  with  an  average  age  of  8-9  months  retained 


PHOSPHORUS  METABOLISM  419 

0.061  gm.  P205,  or  0.027  gm.  P,  on  an  intake  of  0.335  gm.  P205,  or 
0.146  gm.  P  (Netter) ;  with  infants  4-6  months  old  on  artificial  feed- 
ing there  was  a  retention  of  0.015-0.053  gm.  P205,  or  0.0065-0.023 
gm.  P,  on  an  intake  of  0.256-0.273  gm.  P205,  or  0.112-0.119  gm.  P, 
per  day  (Miiller) ;  with  infants  3  and  8^  months  of  age,  and  intakes 
of  0.212  and  0.260  gm.  P205,  or  0.093-0.114  gm.  P,  per  kg.  of  body 
weight,  there  was  retention  of  23.5  and  26.3  percents  of  these 
amounts  (Bruck) ;  the  breast-fed  infant  may  retain  56.5  percent  of 
an  intake  of  0.051  gm.  P205,  or  0.022  gm.  P,  per  kg.  of  body  weight 
per  day  (Tobler  and  Noll).      See  also  Lehmus  (1878). 

PHOSPHORUS  REQUIREMENTS  OF  CATTLE 
Weiske  (1873)  studied  the  metabolism  of  calcium  phosphate 
with  two  5-6-months-old  calves.  One  calf  retained  about  a  half  of 
the  12  grams  of  calcium  phosphate  added  to  the  ration  per  day, 
while  the  other  did  not  retain  any  of  the  added  calcium  phosphate, 
the  difference  in  results  apparently  being  due  to  the  greater 
consumption  of  the  basal  ration  by  the  latter  calf,  the  food  furnish- 
ing the  entire  calcium  and  phosphorus  requirement.  Weiske  consid- 
ered that  16.85  gm.  CaO  and  21.88  gm.  P205  probably  represents 
the  full  daily  requirement  of  the  calves  for  these  elements. 

F.  Soxhlet  (1878),  experimenting  with  3  young  calves  on  a 
milk  diet,  determined  the  average  daily  storage  per  kilogram  of  live 
weight  of  the  several  mineral  constituents  as  follows : 

DAILY  MINERAL  RETENTION  PER  KILOGRAM  OF  LIVE  WEIGHT  OF 
THE  CALF  RECEIVING  MILK  ALONE 

Ash 0.657  gm 53  percent  of  the  food 

P205 0.274  gm 72.5  percent  of  the  food 

CI    0.051  gm 3.8  percent  of  the  food 

CaO     0.286  gm 97.0  percent  of  the  food 

MgO    0.008  gm 30.5  percent  of  the  food 

K20   0.065    gm 20.7  percent  of  the  food 

Na20 0.027  gm 29.1  percent  of  the  food 

Fea03  0.0008  gm 38.1  percent  of  the  food 

J.  Neumann  (1893b,  1894)  found  that  a  calf  weighing  65.63  kg., 
when  fed  on  milk  alone,  stored  both  calcium  and  phosphorus  rapidly. 
On  a  daily  allowance  of  15000  gm.  skim  milk,  and  an  intake  of  24.63 
gm.  CaO,  the  calf  retained  11.77  gm.  CaO.  The  intake  of  P205  was 
30.46  gm.,  and  the  retention  15.55  gm. 

We  are  not  able  to  present  figures  of  any  great  value  as  indi- 
cating the  phosphorus  requirements  of  milch  cows.  On  account  of 
the  nature  of  the  case  especial  value  would  attach  to  observations 
covering  the  whole  period  of  pregnancy  and  lactation. 

Anger  (1898)  made  a  study  of  mineral  metabolism  in  milch 
cows.  The  cows  were  fed  the  following  basal  ration :  50  kg.  fodder 
beets,  6  kg.  straw,  12  kg.  hay  per  1000  kg.    live    weight,    and    in 


420 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


addition  a  supplementary  allowance  of  grain,  mill  feed,  or  commer- 
cial concentrate,  the  amounts  of  the  supplements  being  so  adjusted 
as  to  furnish  the  same  weight  of  protein  in  each  case. 

Anger  concludes  from  this  work  that  the  customary  assumption 
that  ordinary  foods  contain  as  much  mineral  nutriment  as  is  needed 
is  not  justified.  The  intake  and  outgo  of  calcium  and  phosphorus 
are  in  the  following  table. 

CALCIUM  AND  PHOSPHORUS  BALANCES  WITH  MILCH  COWS 
Rations  Varied  from  Period  to  Period — Grams 


CaO 

P2O5 

Week  of 
lacta- 

Milk pro- 
duced daily 

tion 

Kg. 

Intake 

Outgo 

Balance 

Intake 

Outgo 

Balance     i 

20 

9.53 

39.05 

45.38 

-6.33 

66.44 

47.46 

+18.98 

22 

12.05 

41.64 

52.88 

-11.24 

78.72 

79.11 

-0.39    / 

26 

10.54 

55.78 

51.31 

+4.47 

185.25 

143.45 

+41.80 

8.07 

41.86 

51.57 

-9.71 

52.24 

92.34 

-40.10 

34 

6.19 

49.81 

50.20 

-0.39 

59.84 

52.88 

+6.96 

38 

4.71 

62.51 

49.80 

+12.71 

97.27 

98.88 

-1.61 

41 

3.11 

99.07 

95.33 

+3.74 

99.20 

95.25 

+3.95 

44 

2.82 

52.57 

49.34 

+3.23 

76.49 

68.25 

+8.24 

2 

41 

8.12 

39.65 

52.92 

-13.27 

•    48.35 

42.25 

+6.10 

2 

43 

7.81 

62.32 

45.66 

+16.66 

95.91 

84.09 

+11.82    , 

2 

47 

5.92 

95.27 

91.28 

+3.99 

92.19 

85.57 

+6.62 

2 

51 

3.91 

40.40 

36.10 

+4.30 
+6.43 

65.90 

73.90 

-8.00 

2 

55 

1.32 

33.01 

26.58 

54.94 

65.42 

-10.48 

2 

5 

15.48 

37.16 

67.63 

-30.47 

61.11 

85.32 

-24.21 

3 

17 

11.32 

24.75 

30.90 

-6.15 

63.70 

52.81 

+10.89 

3 

20 

10.00 

40.76 

54.17 

+13.41 

155.32 

144.13 

+11.19 

3 

23 

9.16 

33.36 

48.86 

-15.50 

47.14 

78.03 

-30.86 

4 

49 

8.17 

53.98 

52.33 

+1.65 

90.41 

88.66 

+1.75 

4 

56 

7.84 

55.13 

54.91 

+0.22 

90.13 

87.66 

+2.47 

5 

19 

•   13.85 

54.90 

71.89 

—16. 99 

78.90 

93.59 

-14.69 

5 

23 

11.43 

57.46 

60.95 

-3.49 

57.86 

68.91 

—11.05 

5 

26 

12.62 

58.19 

52.14 

+6.05 

80.70 

77.23 

+3.47 

Note:      There  was   in   each  period   a   gain   in   live   weight. 


From  the  work  of  Jordan,  Hart  and  Patten  (1906)  we  take  the 
following  data  which  have  a  bearing  on  the  phosphorus  requirement 
of  milch  cows. 

AVERAGE  DAILY  PHOSPHORUS  REQUIREMENT  OF  MILCH  COWS 


Weight  of 
cow 

Lbs. 

Daily  milk 
produced 

Kilos 

Phosphorus 
fed  daily 

Grams 

Phosphorus 
stored 
daily 
Grams 

Rations 

1100 
966 

12.813 
12.027 

16.715 
16.768 

14.606 

78.7 
83.3 

37.0 
37.0 

77.0 

8.1 
12.9 

17.6 
17.7 

11.6 

Oat  straw  10  lbs.,  wheat  bran  10  lbs.,  hominy  feed 
5  lbs.,  wheat  gluten  1  lb. 

Oat  straw  10  lbs.,  washed   bran  10  lbs.,  corn  germ 
meal  6  lbs.,  rice  meal  3  lbs. 

Oat  straw  10  lbs.,    wheat  bran  10  lbs.,  rice  meal 
7  lbs.,  wheat  gluten  2  lbs. 

In  their  study  of  the  metabolism  of  the  ash  constituents  of 
wheat  bran  Hart,  McCollum  and  Humphrey  have  recorded  balance 
data  on  high-  and  low-phosphorus  rations  with  milch  cows.      These 


PHOSPHORUS  METABOLISM 


421 


rations  furnished  decidedly  more  or  decidedly  less  phosphorus  than 
the  cow  requires,  and  so  do  not  make  possible  a  close  estimate  of 
their  necessities. 

DAILY  PHOSPHORUS  BALANCES  WITH  COWS  ON  HIGH  AND   LOW 
PHOSPHORUS  RATIONS— Grams 


Dec  1-4... 
Dec  19-22. 
Dec  27-28. 
Jan.  6-9  .. 
Jan.  18-19. 
Jan.  28-31. 

Feb.  5 

Feb.  10-13. 


Ration 


High  phosphorus 

Low 

High         " 

Low 

Hig-h  " 

Low 

High         \\ 

Low 


Intake 

Milk 

Total 

P2O5 

P2O5 

outgo 
P2O5 

190.5 

30.4 

181.5 

46.7 

27.6 

67.6 

190.5 

27.6 

175.2 

47.7 

23.4 

60.3 

190.5 

23.5 

155  0 

41.5 

24.1 

55.4 

190.5 

24.7 

65.6 

46.7 

26.4 

58.0 

Balance 
P2O5 


+9.0 
-20.9 
+15.3 
—12.6 
+35.5 
-13.9 
+124.9 
-11.3 


Gouin  and  Andouard  (1907)  state  that  if  the  ration  of  the  calf 
does  not  contain  legumes  and  milk  it  is  desirable  to  add  100  gm. 
of  powdered  bone  to  the  daily  ration  of  a  calf  of  150  kilograms  live 
weight. 

Khuen  (1908)  determined  in  a  7-day  period  that  with  a  cow 
weighing  429.1  kg.  and  producing,  on  the  average,  7014.3  gm.  milk 
per  day,  an  intake  of  61.88  gm.  CaO,  44.04  gm.  MgO  and  84.76  gm. 
P205  resulted  in  a  loss  of  18.24  gm.  CaO,  a  gain  of  0.21  gm.  MgO  and 
a  loss  of  1.76  gm.  P205.  The  milk  contained  16.31  gm.  CaO,  1.97 
gm.  MgO  and  54.60  gm.  P205. 

From  these  data  we  compute  that  in  addition  to  the  amount  of 
phosphorus  in  the  milk  a  cow  must  receive  more  than  70  mg.  P205 
per  kg.  live  weight. 

Henneberg  (Beitrage  zur  Begriindung  usw.,  2  heft,  1864  S.  53.) 
determined  the  maintenance  requirement  of  the  steer,  for  calcium 
and  phosphorus,  per  100  kg.  live  weight,  as  100  gm.  lime  and  50  gm. 
phosphoric  acid.  Kellner  (1907),  in  computing  the  requirements 
of  the  milch  cow,  adds  to  these  figures  three  times  the  lime  and 
phosphoric  acid  content  of  the  20  kg.  of  milk  produced  per  1000  kg. 
of  live  weight  by  milch  cows,  and  so  obtains  as  a  total  200  gm.  of 
lime  and  140  gm.  of  phosphoric  acid  as  the  requirements  of  milch 
cows  per  1000  kg.  live  weight. 

A.  R.  Rose  (1912a)  concludes  that  the  phosphorus  maintenance 
requirement  of  a  milch  cow  is  the  amount  of  phosphorus  eliminated 
in  the  milk  plus  26  milligrams  (P)  per  kilogram  of  body  weight. 

Kellner  (1907)  computes  the  calcium  and  phosphorus  retention 
of  the  growing  calf  as  21  gm.  CaO  and  19  gm.  P,05  per  day  during 
the  first  year,  and  states  that  the  food  should  contain  40-60  gm.  of 
each. 


422 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


PHOSPHORUS  REQUIREMENTS  OF  HORSES 

There  is  in  existence  but  very  little  evidence  of  value  on  the 
phosphorus  requirements  of  horses,  especially  on  the  optimum 
amounts  under  the  various  conditions  of  growth.  At  the  same  time 
it  is  surely  true  that  there  is  no  other  animal  in  which  the  develop- 
ment and  character  of  the  skeletal  tissues  contributes  in  so  promin- 
ent a  way  to  the  economic  value. 

As  is  true  with  other  herbivora,  little  is  to  be  learned  from  the 
quantitative  estimation  of  urinary  phosphorus,  except  in  connection 
with  the  other  factors  of  a  complete  balance  determination.  A  few 
data  on  the  amounts  of  phosphorus  in  the  urine  of  horses,  however, 
are  perhaps  worthy  of  record. 

Salkowski  (1885  and  1904)  found  the  urinary  phosphorus  of  a 
horse  to  be  about  0.0107  gm.  P205  per  100  c.c,  and  0.2199  gm.  in  24 
hours. 

F.  Smith  (1889)  found  in  horse  urine  sometimes  only  a  trace  of 
phosphorus;  at  others  from  0.13  to  9.45  gm.  P205  for  24  hours,  an 
average  amount  for  rest  being  1.3  gm.,  and  for  work  1.897  gm.  P205. 

Liebermann  (1891c)  found  the  phosphorus  content  of  the 
urine  of  8  stallions  to  vary  between  0.004  and  0.021  percent  P205, 
and  the  24-hour  outgo  of  P205  in  the  urine  of  six  stallions  to  vary 
from  0.118  to  0.481  gm. 

Tangl  (1901)  reported  balance  experiments  on  mature  horses 
with  calcium-deficient  rations,  an  abstract  of  the  report  of  which 
we  have  seen  in  Maly's  Jahresbericht.  The  original  work  contains 
nitrogen,  calcium,  magnesium  and  phosphorus  balances.  The 
abstract  referred  to  contains  little  significant  material.  Tangl 
states  the  opinion  that  the  mature  horse  is  able  to  supply  its  calcium 
needs  from  the  same  low-calcium  fodder,  which,  with  cattle,  may 
produce  malnutrition  of  the  bones. 

Tangl  (1902a,  1902b)  studied  mineral  metabolism  in  growing 
horses,  with  rations  of  low  calcium  content.  Two  horses  were 
used  in  these  experiments.  In  the  first  experiment  they  received 
hay  alone ;  in  the  second  oats  and  hay.    (See  table  on  p.  423) . 

The  length  of  the  first  experiment  was  8  days ;  of  the  second  6 
days.      The  percentage  composition  of  the  foods  is  as  follows : 


Experiment  I 

Experiment  II 

Hay 

Hay 

Oats 

N 
P 
Ca 
Mg 

1.366 
0.146 
0.440 
0.137 

1.844 
0.165 
0.410 
0.209 

2.272 
0.406 
0.143 
0.104 

PHOSPHORUS  METABOLISM 


423 


AVERAGE       DAILY      NITROGEN,       CALCIUM,       MAGNESIUM       AND 

PHOSPHORUS  BALANCES  WITH  HORSES  ON  LOW  CALCIUM 

RATIONS— Grams 


Weights 

N 

Ca 

Mg 

P 

Initial 

Final 

Ave.  daily 

change 

Kg. 

Food 

Food 

Food     , 

Food 

Subject 
No. 

Urine 
Feces 

Urine 
Feces 

Urine 
Feces 

Urine 
Feces 

Rations 

Balance 

Balance 

Balance 

Balance 

109.28 

36.38 

11.23 

11.740 

408.0 

70.39 

14.38 

3.43 

0.061 

8  kg.  bay,  19.6  kg.  water 

1 

404.0 

50.70 

21.31 

7.80 

11.830 

-0.50 

-11.81 

+0.69 

+0.09 

-0.151 

109.28 

36.72 

11.33 

11.74 

445 

68.11 

14.11 

3.46 

0.13 

8  kg.  hay,  26.5  kg.  water 

2 

440 

52.91 

21.84 

7.85 

11.85 

-0.63 

-11.74 

+0.77 

+0.02 

-0.24 

183.08 

27.23 

14.86 

24.50 

403.8 

102.16 

8.10 

3.44 

0.79 

5  kg.  hay,  4  kg.  oats, 

1 

406.5 

48.30 

17.23 

30.09 

23.07 

17.3  kg.  water 

+0.45 

+32.62 

+1.90 

+0.33 

+0.64 

183.08 

27.44 

14.92 

24.50 

432.5 

90.65 

7.69 

3.59 

0.47 

5  kg.  hay,  4  kg.  oats, 

2 

436.8 

53.45 

18.66 

10.70 

23.50 

22.0  kg.  water 

+0.72 

+38.98 

+1.09 

+0.63 

+.053 

The  drinking  water  contained  0.0049  percent  Ca  and  0.0013 
percent  Mg. 

From  these  data  we  compute  that  0.27  gm.  N  in  hay,  per  kg. 
body  weight,  is  below  the  maintenance  requirement,  while  0.45  gm., 
in  hay  and  oats,  results  in  marked  storage ;  the  phosphorus  content 
of  the  hay  ration  limited  calcium  storage,  since  with  increased  phos- 
phorus but  decreased  calcium  intake,  in  the  second  experiment,  there 
was  increased  calcium  storage ;  with  an  intake  of  0.063  gm.  Ca  per 
kg.  live  weight  there  was  Ca  storage ;  with  an  intake  of  0.029  gm.  P 
per  kg.  there  was  loss  of  P,  while  with  an  intake  of  0.056  gm.  P  per 
kg.  there  was  storage  of  this  element. 

Alquier  (1905-6)  determined  that  a  working  horse  of  400-450 
kilograms  (880-990  lbs.)  weight  requires  the  equivalent  of  about  35 
grams  daily  of  P2Og  for  maintenance  of  phosphorus  equilibrium.  By 
the  use  of  a  suitable  amount  of  feeding  molasses,  however,  it  was 
found  that  25.5  grams  of  P205  sufficed  for  the  maintenance  of  equi- 
librium, the  increased  usefulness  of  the  phosphorus  being  considered 
to  be  due  to  increased  solubility,  and  therefore  assimilability,  being 
caused  by  the  action  of  the  sodium  and  potassium  salts  occurring 
abundantly  in  the  molasses. 


424 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


PHOSPHORUS  REQUIREMENTS  OF  SWINE 

From  the  composition  of  a  year's  growth  of  a  pig  Kellner 
(1907)  computes,  allowing  three  times  as  much  in  the  food  as  the 
animal  will  store  in  the  body,  that  the  daily  requirement  of  a  pig  is 
12  gm.  each  of  lime  and  phosphoric  acid. 

Hart,  McCollum  and  Fuller  (1909)  expressed  the  opinion  that 
the  daily  phosphorus  supply  for  a  50-lb.  growing  pig  should  be  at 
least  3  gm.,  and  that  a  supply  of  4  to  5  gm.  is  probably  a  safer  quan- 
tity. From  their  experiments  with  low-phosphorus  rations  contain- 
ing 1.12  gm.  of  phosphorus  per  kilogram  of  feed,  and  from  the 
experiments  of  Forbes  and  associates  (1914)  with  similar  rations 
containing  0.92-0.98  gm.  of  phosphorus  per  kilogram  of  feed  it 
appears  to  be  impossible  to  keep  pigs  alive  indefinitely  on  food  so 
poor  in  phosphorus.  Doubt  attaches  to  this  conclusion,  however, 
because  of  the  fact  that  these  rations  were  also  very  low  in  calcium. 

In  the  course  of  his  investigation  of  nuclein  metabolism  in 
swine  Schittenhelm  (1910)  conducted  balance  experiments  which 
indicated  phosphorus  requirements.  A  part  of  his  figures  are  as 
follows : 

AVERAGE  DAILY  NITROGEN  AND  PHOSPHORUS  BALANCES  WITH  A 

YOUNG  PIG— Grams 


Period 

Duration 
in  days 

N 

Food 

Urine 

Feces 

Balance 

P2O5 
Food 
Urine 
Feces 
Balance 

Ration 

1 
2 
3 

5 
3 
3 

8.70 
5.84 
0.265 
+2.595 

9.49 

6.57 
0.682 
+2.238 

8.70 

6.30 

0.193 

+2.207 

3.80 

1.20 

0.39 

+2.21 

4.40 

1.63 

0.695 

+2.075 

3.80 

1.23 

0.191 

+2.379 

1500  c.  c.  milk 

1500  c.  c.  milk.  20  gm.  yeast  nucleinic  acid 

1500  c.  c.  milk 

1 
2 
3 

5 
5 
5 

14.07 

5.20 

0.55 

+8.32 

16.12 

6.38 

0.21 

+9.52 

14.07 

6.40 

0.295 

+7.375 

5.30 

1.07 

0.45 

+3.78 

7.65 

3.32 

0.20 

+4.13 

5.30 

6.'3i 

1500  c.  c.  milk,  300  gm.  wheat  flour 

1500  c.  c.  milk, 

300  gm.  wheat  flour, 

14  gm.  yeast  nucleinic  acid 

1500  c.  c.  milk, 

300  gm.  wheat  flour 

PHOSPHORUS  METABOLISM  425 

In  the  first  experiment  the  pig  weighed  about  15  kg.  and  is 
said  not  to  have  gained  in  weight ;  it  was  8  weeks  old ;  in  the  second 
experiment  the  same  pig  was  used,  at  an  age  of  4  months.  The 
initial  weight  was  25.4  kg.,  and  the  final  weight  31.6  kg.  In  both 
experiments  there  was  a  large  storage  of  nitrogen  and  phosphorus. 
The  energy  value  of  the  ration  in  the  first  experiment  was  deficient. 
In  the  second  experiment  the  increase  in  weight  was  413  gm.  daily, 
with  an  intake  of  about  0.5  gm.  nitrogen,  and  0.2  gm.  P205  per  kg. 
of  body  weight. 

In  balance  experiments  on  young  swine  Weiser  (1912)  showed 
that  calcium  is  the  limiting  factor  in  the  storage  of  phosphorus  on 
a  ration  of  corn  alone.  By  the  addition  of  calcium  carbonate  to  the 
corn  ration  the  phosphorus  balance  changed  from  negative  to  posi- 
tive. It  seems  certain,  therefore,  that  cereal  foods  generally,  at 
least  corn  and  barley,  contain  enough  phosphorus  to  make  storage 
possible  if  the  calcium  is  increased  to  an  adequate  amount.  If,  how- 
ever, the  protein  were  increased  to  such  an  amount  as  would  provide 
for  maximum  growth,  by  the  addition  of  a  more  highly  nitrogenous 
food,  it  may  be  that  the  phosphorus  of  the  ration  would  then  limit 
production,  this  depending  in  large  measure  on  the  phosphorus  con- 
tent of  the  proteid  supplement.  On  rations  of  corn  alone  Weiser 
got  negative  phosphorus  balances  on  intakes  of  0.057,  0.050,  and 
0.049  gm.  phosphorus  per  kg.  live  weight,  and  marked  storage  on  an 
intake  of  0.048,  0.060,  and  0.051  gm.  phosphorus  per  kg.  live  weight 
on  barley  and  starch,  or  on  corn  and  calcium  carbonate,  or  barley 
and  calcium  carbonate. 

From  the  experiments  of  Forbes,  Beegle,  Fritz  and  Mensching 
(table  p.  378)  it  would  appear  that  actual  phosphorus  requirements 
of  growing  swine  are  satisfied  by  corn  and  the  supplementary  foods 
ordinarily  fed  with  corn,  but  the  optimum  allowance  of  phosphorus 
has  not  been  determined.  In  these  experiments  phosphorus  was 
retained  on  an  intake  of  48  mg.  per  kg.  of  live  weight. 

PHOSPHORUS  REQUIREMENTS  OF  SHEEP 

No  very  satisfactory  data  on  this  subject  have  come  to  hand. 
Weiske  (1880)  found  that  a  ration  of  meadow  hay  and  peas 
contained  enough  of  each  of  the  mineral  nutrients  to  provide  for 
liberal  storage  at  all  stages  of  maturity. 

Jordan  (1885-6)  found  that  timothy  hay  and  cottonseed  meal 
would  sustain  large  retention  of  phosphorus,  while  timothy  hay  and 
corn  led  to  moderate  retention  of  phosphorus  by  a  mature  wether. 
On  the  following  page  are  data  from  this  test. 


426 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


AVERAGE  DAILY  PHOSPHORUS  AND  NITROGEN  BALANCES  WITH  A 
MATURE  WETHER  ON  NORMAL  FOODS— Grams 


Nitrogen 

Phosphorus 

Rations 

Food 

Urine 

Feces 

Bal- 
ance 

Food 

Urine 

Feces 

Bal- 
ance 

20.72 

16.78 

5.36 

-1.42 

8.08 

0.00 

7.48 

+0.60 

Timothy  hay  600  gm.; 
cottonseed  meal  200  gm. 

9.02 

4.59 

4.12 

+0.31 

2.72 

0.00 

2.48 

+0.24 

Timothy  hay  600  g-m.; 
corn  meal  200  gm. 

The  first  period  was  of  five  days  duration  and  the  second  of  four 
days.      No  phosphorus  was  found  in  the  urine. 

The  following  figures  from  Weiske's  work  with  sheep  (copied 
from  Wolff)  are  of  some  value  as  indicating  mineral  requirement. 

DAILY  RETENTION  OF  MINERAL  MATTER  BY  SHEEP— Grams 


Age  of  sheep 
Months 

Live  weight 
Lhs. 

Potash 

Soda 

Lime 

Magnesia 

Phosphoric  acid 

5-6 

7-9 

10-12 

13-15 

51 
66 

77 
85 

2.04 
2.89 
3.05 
2.65 

0.84 
1.05 
0.81 
0.72 

1.56 
2.00 
1.81 
2.07 

0.12 
0.32 
0.38 
0.35 

1.09 
1.65 
2.50 
3.14 

As  to  how  many  times  these  amounts  of  the  mineral  nutrients 
the  food  should  contain,  however,  we  are  unable  to  say  with  assur- 
ance. Wolff  states  that  the  food  of  young  animals  reared  artificially 
should  contain  2  to  3  times  as  much  lime  and  phosphoric  acid  as  that 
actually  required  as  nutriment  by  the  animals. 

Kellner  (1907)  computes,  on  the  basis  of  Weiske's  experiments 
with  lambs  4-5  months  old,  that  the  daily  retention  per  50  kg.  of 
live  weight  is  2.5-3.8  gm.  of  lime  and  2.0-4.1  gm.  of  phosphoric  acid. 
Then,  allowing  2-3  times  these  quantities  in  the  food,  he  recom- 
mends 22  gm.  of  lime  and  25  gm.  of  phosphoric  acid  per  100  kg.  live 
weight  per  day. 

PHOSPHORUS  REQUIREMENTS  OF  DOGS 

In  investigations  by  Forster  (1873a),  with  dogs  on  low-ash 
experimental  rations,  one  dog  weighing  about  24  kg.  was  in  phospho- 
rus equilibrium  on  about  0.04  gm.  P205,  or  0.017  gm.  P,  per  kg.  body 
weight,  daily.  This  may  be  considered  as  a  minimum  figure  for 
artificial  conditions. 

Munk  (1894)  showed  that  a  mature  dog  weighing  normally 
17.16  kg.  lost,  per  day,  during  fasting,  4.68  gm.  nitrogen  and  0.98  gm. 
P2Os  in  the  urine,  and  0.158  gm.  nitrogen  and  0.187  gm.  P205  in  the 


PHOSPHORUS  METABOLISM  427 

feces,  a  total  therefore  of  4.838  gm.  nitrogen  and  1.167  gm.  P205 
per  day,  and  0.0282  gm.  nitrogen  and  0.068  gm.  P205,  or  0.030  gm.  P, 
per  kg.  per  day. 

In  metabolism  studies  by  Zadik  (1899),  with  dogs,  there  was, 
during  the  5th  period,  very  nearly  a  condition  of  phosphorus  equi- 
librium. A  dog  weighing  10706  gm.  stored  0.009  gm.  phosphorus 
per  day  in  a  6-day  period  on  a  total  intake  of  0.634  gm.  phosphorus 
per  day,  or  0.059  gm.  per  kg.  live  weight.  The  phosphorus  was 
present  mostly  in  casein.  The  nitrogen  balance  was  negative,  the 
average  daily  loss  being  0.37  gm.  on  an  intake  of  7.25  gm. 

Kornauth  (1900),  in  comparing  the  utilization  of  different  pro- 
teins with  dogs,  obtained  balance  figures  which  show  that,  when  the 
principal  source  of  the  protein  is  meat,  an  intake  of  about  55-60  mg. 
P,  per  kilogram  of  body  weight  per  day,  is  close  to  the  maintenance 
requirement. 

On  a  diet  of  meat,  lard,  crackers  and  bone  ash,  Hawk  and  Gies 
(1904)  maintained  a  dog  weighing  16.96  kg.  for  12  days  on  an  intake 
of  2.456  gm.  of  phosphorus  per  day,  with  a  daily  phosphorus  balance 
of  +0.045  gm.  Of  this  2.456  gm.  phosphorus,  1.779  gm.  was  pres- 
ent as  bone  ash. 

With  another  dog,  on  the  same  ration,  these  authors  fed  the 
animal  1.95  gm.  phosphorus  daily  (apparently  not  0.195  gm.  as 
stated  in  the  original),  of  which  1.423  gm.  was  present  as  bone  ash. 
The  dog  weighed  11.85  kg.,  and  the  daily  phosphorus  balance  was 
—0.030  gm. 

The  apparent  phosphorus  requirement  with  these  dogs  was 
about  0.140-0.170  gram  per  kg.  live  weight,  the  amount  being  due 
in  part  to  the  comparatively  insoluble  form  in  which  most  of  the 
phosphorus  was  fed. 

Heubner  (1909)  determined  that  the  phosphorus  retention  of 
the  normal  growing  dog  was  0.14  gm.  per  kilogram  of  body  weight 
per  day. 

The  lowest  amounts  of  calcium  and  phosphorus  on  which  stor- 
age took  place  in  Biernacki's  experiments  (1909)  were  0.044  gram 
CaO  and  0.248  gram  P205,  or  0.108  gm.  P,  per  kg.  live  weight.  The 
amount  of  phosphorus  stated,  however,  is  considerably  above  the 
maintenance  requirement. 

In  an  investigation  on  the  use  of  organic  and  inorganic  phos- 
phates in  foods,  Lipschutz  (1911b)  determined  that,  in  a  7-day  bal- 
ance experiment,  a  dog  weighing  2450  gm.  at  the  beginning,  and 


428  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

2700  gm.  at  the  end,  with  a  ration  of  rice,  egg  albumin,  sugar,  palmi- 
tin,  and  a  salt  mixture,  including  phosphates,  and  with  a  daily 
phosphorus  intake  of  0.651  gm.,  or  0.252  gm.  per  kg.  live  weight,  the 
retention  was  0.084  gm.,  or  0.033  gm.  per  kg.  live  weight  per  day. 

Kochmann  and  Petzsch  (1911),  in  their  study  of  calcium 
metabolism  in  the  dog,  made  observations  on  phosphorus  metabo- 
lism from  which  it  is  possible  to  estimate  the  phosphorus  require- 
ment. The  dog  in  question  was  mature,  and  weighed  10300-11040 
gm.  in  the  periods  in  which  there  was  a  retention  of  P205.  In  4 
periods  of  4-5  days  each  the  P205  balances,  on  an  intake  of  3.671  gm., 
were  +0.308,  +0.121,  —0.260  and  +0.622.  With  an  intake  of 
2.800  gm.  P205  the  phosphorus  balance  was  negative.  The  phos- 
phorus requirement,  therefore,  appeared  to  be  about  0.114-0.142 
gm.  per  kg.  body  weight. 

From  these  data  it  appears  that  the  fasting  loss  is  about  0.030 
gm.  per  kilogram  (Munk) ;  the  maintenance  requirement  on  an  arti- 
ficial low-ash  diet  about  0.017  gm.  per  kilogram  (Forster) ;  the  main- 
tenance requirement  on  a  more  nearly  normal  diet,  with  the  phos- 
phorus present  mostly  in  casein,  was  0.059  gm.  per  kilogram 
(Zadik) ;  with  a  diet  containing  meat  and  bone  ash  the  requirement 
seemed  to  be  about  0.140-0.170  gm.  per  kilogram  (Hawk  and  Gies) ; 
retention  took  place  on  intakes  of  0.108  gm.  (Biernacki),  and  0.252 
gm.  (Lipschiitz),  while  Heubner  showed  that  the  normal  retention 
is  about  0.140  gm.  per  kilogram  in  a  growing  dog. 

The  data  are  insufficient  for  the  establishment  of  general 
conclusions. 

PHOSPHORUS  REQUIREMENTS  OF  RATS 

From  the  experiments  of  Gregersen  (1911)  we  deduce  the 
observations  that  on  a  nitrogen-free,  phosphorus-free  diet  the  daily 
elimination  of  phosphorus  per  1000  gm.  of  body  weight  is  36.& 
milligrams,  the  eight  figures  averaged  varying  from  28  to  54.5  milli- 
grams. From  an  experiment  by  McCollum  (1909),  we  compute  from 
a  determination  with  a  single  individual  on  a  phosphorus-free  diet, 
that  the  outgo  was  35.0  milligrams  per  1000  gm.  of  body  weight. 
The  experiments  of  neither  author  afford  satisfactory  evidence  for 
judgment  as  to  the  maintenance  or  growth  requirement  on  normal 
foods. 


PHOSPHORUS  METABOLISM 


429 


PHOSPHORUS  METABOLISM  AS  AFFECTED  BY  VARIOUS 

CONDITIONS 

EFFECTS  OF  ALTITUDE  ON  PHOSPHORUS  METABOLISM 

Von  Wendt  (1910)  studied  metabolism  in  man  at  three  differ- 
ent altitudes  in  the  Alps.  Complete  nitrogen  and  mineral  balances 
were  made.      A  portion  of  the  data  are  as  follows : 

DAILY  NITROGEN  AND  MINERAL  BALANCES    WITH    A    MAN     AT 
DIFFERENT  ALTITUDES  IN  THE  ALPS— Grams 


Day 

Altitude 
Meters 

N 

Food 

Urine 

Feces 

Balance 

S 

Food 

Urine 

Feces 

Balance 

Fe 

Food 

J  Urine 

1  Feces 

Balance 

Ca 

Food 

Urine 

Feces 

Balance 

P 

Food 

Urine 

Feces 

Balance 

1 
2 
3 
4 

3000 
3000 
3000 
3000 

22.58 

13.64 

1.42 

+7.52 

22.58 

16.02 

1.42 

+5.14 

22.58 

18.23 

1.42 

+2.93 

22.58 

15.70 

1.42 

+5.46 

1.706 

1.062 

0.343 

+0.301 

1.706 

1.343 

0.343 

+0.020 

1.106 

0.963 

0.343 

+0.400 

1.706 

1.123 

0.343 

+0.240 

0.156 

0.101 

+0.055 

0.156 

0.101 

+0.055 

0.156 

0.101 

+0.055 

0.156 

0.101 

+0.055 

0.160 

0.177 

0.200 

-0.217 

0.160 

0.197 

0.200 

-0.237 

0.160 

0.140 

0.200 

-0.180 

0.160 

0.157 

0.200 

-0.197 

0.861 

0.661 

0.197 

+0.003 

0.861 

0.774 

0.197 

-0.110 

0.861 

0.766 

0.197 

-O.102 

0.861 

0.828 

0.197 

-0.164 

1 

2 
3 

4 

4560 
4560 
4560 
4560 

22.58 

14.58 

1.22 

+6.78 

22.58 

14.38 

1.22 

+6.98 

22.58 

15.00 

1.22 

+6.36 

3.30 

9.97 

1.22 

-7.89 

1.706 

0.892 

0.282 

+0.532 

1.706 

0.928 

0.282 

+0.496 

1.706 

1.086 

0.282 

+0.338 

0.435 

0.672 

0.282 

-0.519 

0.156 

0.140 

+0.016 

0.156 

0.140 

+0.016 

0.156 

0.140 

+0.016 

0.003 

0.140 

-0.137 

0.160 

0.193 

0.239 

-0.272 

0.160 

0.137 

0.239 

-0.216 

0.160 

0.143 

0.239 

—0.222 

0.083 

0.124 

0.239 

-0.280 

0.861 

0.585 

0.184 

+0.092 

0.861 

0.658 

0.184 

+0.019 

0.861 

0.555 

0.184 

+0.012 

0.284 

0.508 

0.184 

-0.408 

1 
2 
3 

3000 
3000 
3000 

22.58 

14.25 

1.06 

+7.27 

22.58 

15.88 

1.06 

+5.64 

3.30 

11.90 

1.06 

-8.86 

1.706 

0.950 

0.331 

+0.425 

1.706 

1.198 

0.331 

+0.177 

0.435 

1.112 

0.331 

-1.008 

0.156 

0.096 

+0.060 

0.156 

0.096 

+0.060 

0.003 

0.096 

-0.093 

0.160 

0.191 

0.216 

-0.247 

0.160 

0.176 

0.216 

-0.232 

0.083 

0.145 

0.216 

-0.278 

0.861 

1.057 

0.241 

-0.437 

0.861 

1.140 

0.241 

-0.520 

0.284 

0.629 

0.241 

-0.586 

430  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

The  first  and  third  series  of  balances  were  at  an  altitude  of  3000 
meters,  and  the  second  at  4560  meters.  The  nitrogen  balance  was 
positive  at  all  times,  except  on  the  two  days  of  low  nitrogen  intake. 
Reckoned  as  muscle  substance,  more  than  100  gm.  per  day  was 
formed.  The  parallel  sulphur  balances  showed  that  the  nitrogen 
storage  was  protein  synthesis.  The  phosphorus  balance  was  posi- 
tive only  during  the  second  period,  at  the  higher  altitude.  Von 
Wendt  believes,  from  the  relative  amounts  of  nitrogen,  sulphur, 
phosphorus  and  iron  stored,  that  during  the  first  and  third  periods, 
at  the  lower  altitude,  there  was  much  haemoglobin  synthesized, 
while  synthesis  of  muscle  substance  predominated  during  the  second 
period  at  the  higher  altitude. 

THE  INFLUENCE  OF  AMOUNT  OF  FOOD  ON  PHOSPHORUS 
METABOLISM 

The  plane  of  nutrition  on  which  an  animal  is  living,  as  deter- 
mined by  the  nutrients  available,  naturally  involves  phosphorus  as 
well  as  the  other  essential  constituents.  Thus  Pettenkofer  and  Voit 
(1866)  showed  that  on  a  medium  diet  the  phosphorus  in  the  urine 
was  one-third  more  than  during  fast,  the  subject  being  at  rest  in 
both  cases,  and  Forster  (1873a),  experimenting  with  foods  poor  in 
mineral  matter,  showed  that  the  less  the  quantity  of  the  salt-poor 
foods  ingested  the  greater  was  the  loss  of  phosphorus,  due  to  tissue 
katabolism,  from  the  body.  An  observation  of  similar  portent  was 
made  by  Weiske  (1894),  who  found  that  in  the  less  well-developed 
rabbits  of  the  same  age  and  kind  the  bones  and  also,  to  a  less  extent, 
the  teeth,  contained  subnormal  percentages  of  mineral  matter. 

Gevaerts  (1901),  in  a  study  of  feeding  white  rats  on  phospho- 
rus-free foods,  submits  data  which  we  have  condensed  as  in  the  table 
on  p.  431. 

From  these  data  we  see  that  in  the  white  rat  the  phosphorus 
excretion  in  the  urine  from  a  phosphorus-free  diet  is  very  much  less 
in  amount  than  that  present  in  the  urine  during  starvation ;  and  also 
that  on  a  ration  of  sucrose  and  edestin,  or  on  sucrose  and  ovalbumin, 
there  is  much  less  phosphorus  in  the  urine  than  on  a  ration  of 
sucrose  alone,  the  edestin  and  ovalbumin  naturally  being  more 
efficient  than  the  sucrose  to  spare  the  phosphorus-containing  pro- 
teins of  the  body. 

Sherman  (1902)  showed,  in  experiments  with  human  subjects, 
that  nitrogen,  sulphur  and  phosphorus  balances  all  reflected  consist- 
ently, by  change  of  sign,  the  change  from  restricted  to  liberal  diet, 
or  the  reverse.  (Exp.  7-10  p.  432.)  The  diet  was  crackers,  butter 
and  milk.  It  is  elsewhere  shown  that  nitrogen,  sulphur  and  phos- 
phorus balances  need  not,  of  necessity,  show  interdependence  in 
their  variations. 


PHOSPHORUS  METABOLISM 


431 


AVERAGE  DAILY  EXCRETION  OF  PHOSPHORUS  IN  THE  URINE  OF 
WHITE  RATS  ON  PHOSPHORUS-FREE  DIETS— Grams 


Experiment, 

period  and 

days 

Ration 

Initial 

live 
weight 

Gain  or 
loss  in 

weight 

Loss  in 
phosphorus 

1  A— 3 

210 

198 

-8.3 

-7.7 

0.0093 

IB— 3.. 

0.0141 

2  A— 2             

182 
159 
147 
145 

-11.5 
-4.0 
-2.0 

+7.0 

0.0060 

2  B— 3 

0.00085 

2c    1.. 

0.0050 

2D— 1 

0.0033 

3  A— 2. 

181 

158 
155 
146 

-11.5 
—1.0 
-3.0 

+8.0 

0.0060 

3B— 3 

0.0010 

3  c— 3    

0.0040 

3D— 1 

0.0038 

4  A— 2 

202 
183 
168 
160 

-9.5 
-5.0 
-2.7 

+8.0 

0.0060 

4B— 3 

0.0010 

4  C— 3 

0.0031 

4  D— 1.. . . 

0.0017 

5  A— 6..                      

255 
180 
153 
152 
148 
147 
128 
128 

—1.7 
—27.0 
-1.0 
—4.0 
—1.0 
-6.3 
0.0 
+1.0 

0.0041 

5  b— 1              

0.0124 

5C— 1 

0.0019 

5D— 1 

0.0008 

5E— 1.. 

0.0013 

5F— 3... 

5G— 2 

BH-2 

0.0018 

6  A— 1 

240 
213 
195 
178 
173 

-27.0 
-6.0 
-5.7 
-2.5 

+4.0 

0.0124 

6B— 3.. 

0.0005 

6D— 2. 

6E— 2 

0.0015 

7  A— 1 

280 
252 

-28.0 
-0.7 

0.0124 

7B— 3 

0.0029 

8A-L 

290 
268 

—22.0 
-2.5 

0.0072 

8B— 2 

0.0036 

432 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


AVERAGE      DAILY      NITROGEN,      SULPHUR      AND      PHOSPHORUS 

BALANCES  WITH  A  MAN  ON  A  DIET  OF  CRACKERS, 

BUTTER  AND  MILK— Grams 


N 

S 

P 

Food 

Experi- 

Dura- 

Fuel 

tion  in 
days 

value 
Cals. 

Pood 
Urine 

Food 
Urine 

Food 
Urine 

No. 

Remarks 

Feces 

Feces 

Feces 

Crack- 

Milk 

Butter 

Balance 

Balance 

Balance 

ers 

15.82 

1.12 

2.29 

14.31 

0.95 

1.41 

0.74 

0.06 

0.78 

1 

4 

2908 

+0.77 

15.82 
14.67 
0.67 

+0.11 

1.12 
0.97 
0.06 

+0.10 

2.29 

1.48 
0.73 

300 

2040 

40 

Fore-period 

2 

4 

2901 

+0.48 

15.82 
14.16 
0.70 

+0.09 

1.12 
0.93 
0.06 

+0.08 

2.29 
1.42 
0.78 

300 

2040 

40 

Loss  of  sleep 

3 

4 

2913 

+0.96 

+0.13 

+0.09 

300 

2040 

40 

After-period 

12.05 

0.94 

1.40 

10.43 

0.72 

1.03 

0.74 

0.09 

0.44 

4 

4 

2082 

+0.88 

18.52 
16.50 
0.87 

+0.13 

1.44 
1.12 

0.08 

-0.07 

3.07 
1.74 
1.10 

405 

1000 

60 

Low  protein 

5 

4 

2607 

+1.15 

+0.24 

+0.23 

120 

3060 

0 

High  protein 

9.76 

0.69 

1.60 

10.94 

0.75 

1.12 

0.27 

0.02 

0.37 

Intermediate 

6 

4 

1555 

-1.45 

-0.08 

+0.11 

120 

1530 

0 

period 

10.11 

0.70 

1.58 

' 

11.51 

0.83 

1.00 

0.45 

0.04 

0.60 

■       7 
j 

5 

1660 

-1.85 

20.22 

15.52 

1.04 

-0.17 

1.40 
1.06 
0.09 

-0.02 

3.16 
1.49 
1.42 

150 

1500 

0 

Food  restricted 
Twice  as  much 

8 
1 

5 

3336 

+3.66 

10.11 
12.87 
0.43 

+0.25 

0.70 
0.88 
0.04 

+0.25 

1.58 
1.32 
0.50 

300 

3000 

0 

as  above 

9 

5 

1656 

-3.19 

-0.22 

-0.24 

150 

1500 

0 

Food  restricted 

f 

20.70 

16.58 

1.05 

1.36 
1.21 
0.10 

3.26 
1.49 
1.37 

Twice  as  much 

10 

3 

3329 

+3.07 

+0.05 

+0.40 

300 

3000 

0 

as  above 

Marked  loss  of  sleep  for  3  successive  nights  resulted  in  a  small 
increase  in  the  excretion  of  nitrogen,  sulphur  and  phosphorus,  the 
proportions  not  being  markedly  abnormal.  The  increased  elimina- 
tion due  to  loss  of  sleep  did  not  appear  until  the  third  day,  while 
changes  resulting  from  alteration  of  the  diet  were  always  percept- 
ible on  the  first  day.  The  elimination  of  phosphorus  by  the  intes- 
tine was  large  and  variable. 


PHOSPHORUS  METABOLISM 


433 


Kaufmann  and  Mohr  (1903)  studied  metabolism  during  over- 
feeding1, following  an  extended  period  of  underfeeding,  with  two 
adult  human  beings.  Both  subjects  were  in  bodily  health  during 
this  experiment  on  forced  feeding.  The  average  daily  nitrogen, 
phosphorus  and  calcium  storage  in  grams  was  as  indicated  below : 


Days 

N 

P2O5 

CaO 

Body 

Subject 

Intake 

Retention 

Intake 

Retention 

Intake 

Retention 

weight 
Kg-. 

1 

2 

7 
10 

18.38 
16.45 

4.90 
5.67 

6.20 
5.36 

1.35 
1.37 

6.07 
4.46 

2.34 
2.10 

62.0 
57.7 

The  amount  of  retention  is  much  influenced  by  the  state  of 
impoverishment  existing,  and  the  nutriment  available. 

Hawk  (1903)  compared  phosphorus  balance  on  moderate  and 
excessive  protein  intake.  With  the  excessive  ingestion  of  protein 
there  was  a  negative  phosphorus  balance,  while  with  less  protein  in 
the  fore-  and  after-periods  there  were  positive  balances.  The  inges- 
tion of  4.96  gm.  per  day  of  phosphorus  (stated  as  P205)  provided  for 
storage  of  this  element  in  men  of  56.2-60  kg.  body  weight. 

In  this  experiment  the  sudden  marked  increase  in  phosphorus 
intake  coincident  with  excessive  protein  ingestion,  so  greatly 
increased  the  urinary  phosphorus  excretion  as  to  change  a  positive 
to  a  negative  balance. 


DAILY  PHOSPHORUS  BALANCES   ON  ADULT  MEN  WITH  VARYING 
AMOUNTS  OF  PROTEIN  IN  THE  FOOD— Grams 


Subject 

Period 

Leng-th 
of  period 
in  days 

Pood 
P2O5 

Urine 
P2O5 

Feces 
P2O5 

Balance 
P2O5 

Rations 

H 
H 
H 

I 

II 
III 

4 
1 
4 

4.96 
5.82 
4.96 

2.58 
3.28 
2.57 

2.07 
3.01 
1.92 

+0.32 
-0.47 
+0.47 

300  gm.  crackers, 
60  gm.  butter, 
1650  gm.  milk. 

225  gm.  beef, 

250  gm.  crackers, 

60  gm.  butter,  1375  gm.milk 

300  gm.  crackers, 
60  gm.  butter, 
1650  gm.  milk. 

R 
R 
R 

I 
II 
III 

4 
1 
4 

4,96 
5.82 
4.96 

3.15 
3.62 

2.78 

1.52 
3.45 
1.89 

+0.29 
—1.25 
+0.29 

Same  as  Period  I  above. 
Same  as  Period  II  above. 
Same  as  Period  III  above. 

Initial  weight   of  H — 56.2   kg.,   of  R — 60   kg. 


434 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


C.  Tigerstedt  (1904)  reported  results  on  nitrogen  and  phospho- 
rus balance  experiments  with  a  young  man  20  years  old  on  rations 
varying  in  phosphorus  content.  Daily  balances,  as  stated  by  the 
author,  we  have  computed  to  average  daily  balances  per  period.  A 
part  of  the  results  are  as  follows : 

AVERAGE  DAILY  PHOSPHORUS  BALANCES  WITH  A  MAN  TWENTY 
YEARS  OLD  ON  DIETS  OF  DIFFERENT  PHOSPHORUS  CONTENT 

Grams 


Nitrogen 

Phosphorus 

Days 

Balance 

Food 

Urine 

Feces 

Balance 

Daily  rations 

1 

2 

3-14 
15-17 

18-22 

23-25 

-13.06 

-7.36 

+1.23 
-2.43 

+2.91 

-0.14 

0.027 

traces 

2.545 
2.515 

3.056 

1.757 

0.649 

0.730 

1.333 
1.506 

1.612 

0.956 

(  mean 
0.134 

1.102 
1.291 

1.253 

0.935 

-0.756 

—0.864 

+0.200 
-0.282 

+0.191 

-0.134 

100  gm.  meal,  18  grin,  sugar,  207  gm.  sago  gruel, 
3  gm.  salt. 

70  gm.  sugar,  270  gm.  jelly,  120  gm.  potato,  80  gm. 
apple. 

About  70  gm.  butter,  750  c.c  milk,  150  gm.  bread, 
430  gm.  potatoes,  150  gm.  roast  meat,  150  gm. 
groats. 

Same  as  above  plus  additional  milk  and  a  casein 
preparation,  and  on  days  19-22, 160  gm.  zweiback. 

First  two  days,  275  gm.  groats,  180  gm.  rye  bread, 
100  gm.  butter;  third  day,    200  gm.   groats,  140 
gm.  rye  bread,  80  gm.  butter. 

Weight   of  subject,    62   kg. 

The  first  two  days'  rations  were  very  low  in  both  nitrogen  and 
phosphorus.  The  results  of  these  two  days  show  that  the  phospho- 
rus arising  from  the  mucous  coat  of  the  intestine  and  from  the  intes- 
tinal juices,  which  appears  in  the  feces,  is  about  0.134  gm.  daily,  but 
this  amount  may  vary  with  the  amount  and  kind  of  food.  This  phos- 
phorus is  less  in  amount  when  food  is  given  which  is  poor  in  both 
nitrogen  and  phosphorus  than  in  starvation,  since  such  foods  serve 
to  protect  body  proteins,  etc.,  from  katabolism. 

The  negative  balance  on  days  15-17  was  due  principally  to 
imperfect  absorption  from  the  intestine. 

The  last  ration  was  a  vegetable  dietary,  except  for  the  butter. 
The  negative  nitrogen  balance  and  the  low  calcium  content  of  the 
cereal  diet  were  both  unfavorable  to  phosphorus  retention. 

From  the  work  of  Liithje  and  Berger  (1904)  (see  p.  218)  we 
would  infer  that  calcium  and  phosphorus  storage  might  take  place 
as  rapidly  in  a  normal  man,  who  had  never  been  ill,  as  in  a  typhus 
convalescent;  but  the  nitrogen  retention  seems  to  be  much  more 
marked  in  the  convalescent,  in  a  greatly  reduced  state,  than  it  is  in 
either  the  well-nourished  convalescent  or  the  normal  cases.  The 
maximum  phosphorus  retention  with  a  convalescent  was  2.832  gm. 
P205,  and  in  a  normal  case  3.381  gm.  P205  daily,  in  each  case  for  10 
days. 


PHOSPHORUS  METABOLISM 


435 


Sweet  and  Levene  (1907)  made  observations  on  nuclein  metabo- 
lism in  a  dog  with  an  Eck  fistula  which  show  the  same  ability  of  the 
organism  to  adapt  itself  to  different  planes  of  nuclein  metabolism 
that  has  frequently  been  noted  with  regard  to  total  nitrogen  and 
phosphorus. 

Vozarik  (1909)  studied  the  effect  of  variation  in  the  amount  of 
protein  in  the  diet  on  the  acidity,  ammonia,  phosphorus  and  total 
nitrogen  in  the  urine  of  children.  Increase  of  protein  increased  the 
outgo  of  all  these  constituents,  both  per  day  and  per  c.c.  of  urine. 

Biernacki  (1909)  sought  to  determine  with  dogs  the  effects 
of  "supernutrition"  on  mineral  metabolism.  The  foods  added  to 
the  standard  diets  were  butter,  sugar  and  eggs.  In  each  case  the 
animal,  though  previously  storing  phosphorus,  accomplished  an 
increased  storage  through  the  ingestion  of  excessive  amounts  of 
these  foodstuffs.  Of  calcium  also  there  was  in  each  case  an 
increased  storage,  or  decreased  loss,  when  the  supplementary  foods 
were  given. 

The  conduct  of  mineral  metabolism  experiments  on  a  really 
satisfactory  basis  requires  that  there  be  intermediate  periods  of 
sufficient  length  to  allow  the  effects  of  previous  feeding  to  disappear. 

Numerical  data  from  this  study  are  below. 

DAILY   NITROGEN,    CALCIUM   AND   PHOSPHORUS    BALANCES    WTH 

DOGS— Grams 


Exper- 
iment 
No. 

Periods 

Change 

in  weight 

of  dog 

Nitrogen 
balance 

CaO 

intake 

CaO 

balance 

P2O5 
intake 

P2O5 
balance 

Diet 

I 

Fore-period 
8  days 

Fat  period 
8  days 

After-period 
8  days 

5700 
-250 

5450 
+170 

5620 
-210 

-0.0264 
+0.5232 
+0.2886 

0.0915 
0.2385 
0.0915 

-0.0019 
+0.1673 
+0.0261 

1.4127 
1.7007 
1.4127 

+0.3527 
+0.8059 
+0.4547 

Meat; 
rice 

Meat; 

rice; 

butter 

Meat; 
rice 

II 

Fore-period 
6  days 

Sugar  period 
6  days 

After-period 
6  days 

5290 
+110 

5400 
+380 

5780 
-160 

+0.8316 
+1.5553 
+0.4855 

0.0927 
0.0927 
0.0927 

-0.0328 
-0.0159 
-0.0301 

1.4776 
1.4776 
1.4776 

+0.5297 
+0.6736 
+0.4845 

Meat; 
rice 

Meat; 
rice; 

sugar 

Meat; 
rice 

in 

Fore-period 
6  days 

Fat  period 
6  days 

After-period 
6  days 

Protein  period 
6  days 

5620 
-120 

5500 
+290 

5790 
-50 

5740 
+130 

-0.1618 
+0.6057 
+0.5142 
+1.6857 

0.1289 
0.2737 
0.1289 
0.1475 

-0.0090 
+0.1147 
-0.0092 
-0.0818 

1.6643 
1.9523 
1.6643 
1.5778 

+0.5540 
+0.5769 
+0.2515 
+0.6692 

Milk; 
rice 

Milk; 

rice; 

butter 

Milk; 
rice 

Milk; 

rice; 

eggs' 

436 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


The  increased  nitrogen  storage  resulting  from  the  addition  of 
butter  and  sugar  indicates  that  these  foods  were  able  to  replace 
a  portion  of  the  protein  previously  used  for  energy  production,  thus 
allowing  its  participation,  along  with  calcium  and  phosphorus,  in 
synthesis  of  tissue. 

Heubner  (1909)  conducted  an  experiment  with  young  dogs  to 
study  the  effects  of  phosphorus  starvation.  He  used  6  pups  taken 
at  weaning  time,  when  38  days  old,  and  fed  them  various  rations  for 
7  weeks,  after  which  a  part  were  killed,  and  their  tissues  examined. 
Two  were  fed  on  a  low-phosphorus  ration  composed  of  egg-white, 
rice,  sugar,  palmin,  ferric  saccharate,  and  sodium,  potassium, 
calcium  and  magnesium  chlorides. 

Three  others  received  a  similar  ration  containing  casein  from 
cow's  milk,  and  with  a  part  of  the  alkali  chlorides  replaced  by 
diphosphates,  so  that  the  food  contained  one  percent  more  phospho- 
rus than  the  above.  The  sixth  dog  had  bread,  milk,  meat,  potato, 
rice  and  fat. 

The  phosphorus-starved  dogs  were  undersized,  their  bones 
became  curved,  and  the  abnormality  of  their  diet  was  in  many  ways 
outwardly  evident.  An  examination  of  the  bones  seemed  to  show 
decided  differences  in  appearance  from  those  of  cases  where 
calcium  starvation  was  known  to  prevail. 

The  feces  of  these  dogs  did  not  contain  phosphorus  in  amounts 
corresponding  with  the  food ;  in  fact,  the  phosphorus  content  of  the 
dry  feces  was  somewhat  nearly  the  same  throughout,  and  apparent- 
ly not  related  to  the  food.  The  phosphorus  content  of  the  urine, 
however,  varied  greatly,  and  in  accord  with  the  amount  in  the  food. 
Data  from  this  work  are  below. 

DATA    SHOWING   PHOSPHORUS   STORAGE   IN   YOUNG   DOGS 


Weight 

Phosphorus 

Phosphorus  storage 

No.  of  dog 

Age  of  dog 
in  weeks 

in  urine 

per  day,  per  kg. 

Nutrition 

Kg. 

Percent 

Grams 

1 

5 

1.13 

0.38 

Mother's  milk 

6 

5 

1.00 

0.51 

Mother's  milk 

3 

7 

1.60 

0.029 

o.'ii 

Cow's  milk 

3 

10.5 

2.80 

0.039 

(0.026) 

Mixed  diet 

3 

17 

4.80 

0.052 

0.037 

Mixed  diet 

1 

7 

1.32 

0.39 

0.13 

P  in  excess 

1 

10.5 

1.80 

0.17 

0.19 

P  in  excess 

6 

11 

2.20 

0.17 

0.23 

P  in  excess 

2 

7 

1.23 

0.0024 

0.008 

P  hunger 

2 

10.5 

1.70 

0.001 

0.015 

P  hunger 

5 

11 

1.90 

0.0007 

0.016 

P  hunger 

Kochmann  (1911)  studied  calcium  metabolism,  as  affected  by 
other  food  constituents,  with  three  grown  dogs  fed  on  dog  biscuit 
and  water.  Below  are  his  figures  on  phosphorus  balance  for  Dog 
III. 


PHOSPHORUS  METABOLISM 


437 


Nine  out  of  twelve  of  the  periods  were  5  days  in  length.  That 
5  days  is  too  short  a  period  for  mineral  metabolism  studies  is  mani- 
fest as  we  compare  periods  2,  3  and  4,  the  intake  being  the  same, 
and  periods  8-12,  the  intake  in  these  also  being  the  same. 

In  the  urinary  figures  there  is  some  evidence  of  delayed  outgo 
as  the  intake  changed;  and  in  the  two  series  above  mentioned  the 
change  in  balance  is  due  principally  to  variation  in  feces  phosphorus, 
the  urinary  phosphorus  remaining  much  more  nearly  constant.  The 
change  in  feces  phosphorus,  especially  its  progressive  decrease  in 
periods  9, 10, 11  and  12,  was  not  interpreted  by  the  author. 

It  is  of  interest  to  note  that  with  each  of  the  three  increases  in 
intake  there  was  a  degree  of  improvement  in  retention  which  the 
animal  was  unable  to  sustain,  since  in  the  next  period  after  the  one 
in  which  the  intake  was  increased  there  was  in  each  case  a  decreased 
retention. 

AVERAGE    DAILY    PHOSPHORUS    (P205)    METABOLISM    OF    A    DOG 
RECEIVING  VARIOUS  AMOUNTS  OF  FOOD— Grams 


Length 

Weight 

Intake  of 

Intake 

Urine 

Feces 

Balance 

Period 

of  period 
in  days 

of  dog 

dog  biscuit 

P2O5 

P2O5 

P2O5 

P2O5 

1 

4 

6750 

100 

2.191 

1.159 

1.326 

-0.293 

2 

5 

6600 

150 

3.286 

1.200 

1.937 

+0.149 

3 

3 

6450 

150 

3.286 

1.243 

2.104 

-0.061 

4 

5 

6280 

150 

3.286 

1.212 

2.262 

-0.188 

5 

5 

6260 

175 

3.833 

1.219 

2.038 

- 

1-0.576 

6 

5 

6320 

175 

3.834 

1.305 

2.502 

0.027 

7 

6 

6355 

200 

4.381 

1.250 

2.251 

-0.880 

1          8 

5 

6420 

200 

3.994 

1.300 

2.467 

-0.227 

9 

5 

6460 

200 

3.994 

1.078 

2.908 

-0.012 

10 

5 

6300 

200 

3.994 

1.056 

2.815 

-0.123 

1        n 

5 

6350 

200 

3.994 

1.016 

2.190 

- 

-0.788 

12 

5 

6450 

200 

3.994 

1.085 

2.020 

- 

-0.889 

Summary.  Balance  experiments  show  us  that  increased  intake 
of  phosphorus  usually  causes  increased  elimination  in  the  urine 
with  carnivora,  in  the  feces  with  herbivora,  and  according  to  the 
nature  of  the  diet  with  omnivora,  the  increased  outgo  being  accom- 
panied by  increased  retention  or  production  at  such  times  as  so 
determined  by  the  general  state  of  nutrition.  Marked  increase  in 
intake,  incidental  to  excessive  protein  ingestion,  is  sometimes  accom- 
panied by  decreased  retention;  but  this  is  not  the  usual  course  of 
metabolism. 

During  deficient  intake  of  phosphorus,  or  of  food  in  general, 
the  consumption  of  phosphorus-free  food  may  decrease  the  loss  of 
phosphorus,  through  the  protection  afforded  the  body  tissues  from 
katabolism. 


438  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Even  in  a  normal  state  of  nutrition,  grown  animals  are  able  to 
increase  their  bodily  reserves  of  phosphorus  to  a  considerable 
extent;  and  after  sickness  the  retention  may  be  very  extensive, 
though  apparently  not  of  a  different  order  from  that  characteristic 
of  normal  growth. 

As  with  other  nutrients,  the  retention  of  phosphorus  is  not 
directly  as  the  intake,  but,  other  conditions  being  favorable,  as  the 
amount  of  intake  in  excess  of  the  maintenance  requirement.  After 
a  certain  point  is  reached,  however,  the  rate  of  increased  retention 
coincident  with  increased  intake,  begins  to  fall  off,  and,  with  indefin- 
itely continued  increase  of  intake,  a  point  is  reached  where  further 
increase  does  not  lead  to  further  storage,  but  only  to  elimination. 

Animals  have  a  capacity  to  adapt  their  phosphorus  exchanges 
to  different  planes  of  metabolism,  as  with  other  nutrients,  nitrogen 
in  particular. 

PHOSPHORUS  METABOLISM   DURING   FAST 

In  fasting  there  is  a  very  considerable  loss  of  phosphorus  from 
the  body,  due  to  the  katabolism  of  the  phosphorus-containing  pro- 
teins which  are  consumed  for  energy  production,  a  much  greater 
loss,  naturally,  than  where  foods  are  used  which  are  poor  in  phos- 
phorus. In  this  latter  case  phosphorus-containing  tissues  are 
protected,  and  a  portion  of  the  katabolized  phosphorus  may  be 
retained  and  utilized  along  with  nutrients  supplied  by  the  food. 
Considerable  attention  has  also  been  given  the  possibility  that  the 
bones  lose  phosphorus  during  starvation. 

I.  Munk  (1887)  announced  conclusions  reached  in  the  course  of 
fasting  experiments  with  Cetti  as  follows :  In  fasting,  the  phospho- 
rus of  the  urine  is  united  partly  with  potassium,  and  partly  with 
calcium  and  magnesium.  The  ratio  of  phosphorus  to  nitrogen  in 
the  urine  is  about  1 :4.5  during  fast,  while  in  the  soft  parts  of  the 
body  the  ratio  is  about  1 :7.  The  increase  of  phosphorus  outgo  is 
both  absolute  and  relative  to  nitrogen.  The  calcium  excretion  is  3 
or  4  times  that  amount  which  would  correspond  to  the  break-down 
of  flesh.  Magnesium  excretion  also  is  much  greater  than  as  if  derived 
from  the  flesh,  but  during  fast  the  calcium  exceeds  the  magnesium. 
From  these  facts  he  concluded  that  the  bones  contribute  to  the  phos- 
phorus loss  during  starvation. 

F.  A.  Falck  (1875),  in  studying  hunger  metabolism  in  a  dog, 
determined  that  a  dog  weighing  21210  grams  lost  in  body  weight 
10380  grams,  and  in  phosphorus  31.7  gm.  P205,  equivalent  to  5706 
gm.  fat-free  fresh  dog-flesh,  while  starving  to  death  in  61*4  days. 


PHOSPHORUS  METABOLISM 


439 


Luciani  (1889)  made  a  general  physiological  study  of  fasting, 
with  Succi  as  a  subject.  In  1889  he  reported  results  from  a  30-day 
fast  from  which  it  would  appear,  in  harmony  with  the  conclusions 
of  Munk,  that  the  nitrogen  and  the  phosphorus  of  the  urine  have 
in  part  an  independent  origin  early  in  the  fasting  period,  as  well  as 
late,  but  most  markedly  so  after  a  considerable  duration  of  the  fast. 
A  portion  of  his  figures  are  as  follows : 

DAILY  PHOSPHORUS  ELIMINATION  IN  THE  URINE  DURING  FAST 


Day  of 

;    fast 

P 

Grams 

N/P 

Day  of 
fast 

P 

Grams 

N/P 

Day  of 

fast 

P 
Grams 

N/P 

1 

0.842 

16.3 

11' 

0.620 

12.7 

21 

0.325 

11.9 

2 

0.898 

12.2 

12 

0.441 

16.2 

22 

0.313 

10.2 

3 

0.920 

15.0 

13 

0.137 

25.6 

23 

0.459 

10.4 

4 

0.925 

13.8 

14 

0.434 

12.2 

24 

0.344 

16.1 

5 

1.031 

12.4 

15 

0.449 

11.4 

25 

0.257 

27.4 

6 

0.932 

13.0 

16 

0.468 

11.7 

26 

0.341 

17.7 

7 

0.814 

11.5 

17 

0.531 

11.6 

27 

0.375 

14.3 

8 

0.691 

12.1 

18 

0.439 

12.3      ' 

28 

0.349 

16.0 

9 

0.593 

13.1 

19 

0.415 

12.3 

29 

0.345 

11.8 

10 

0.544 

12.4 

20 

0.382 

11.5 

30 

0.444 

14.9 

C.  Lehmann,  Miiller,  Munk,  Senator  and  Zuntz  (1893)  reported  on 
fasting  experiments  with  Cetti,  the  professional  faster,  and  Breit- 
haupt,  a  cobbler.  Both  drank  water  at  will,  Cetti  took  no  exercise ; 
Breithaupt  assisted  in  laboratory  work. 

DAILY  NITROGEN  AND  PHOSPHORUS  EXCRETION  IN  THE  URINE  BY 
CETTI  AND  BREITHAUPT  DURING  FAST— Grams 


Cetti 

Breithaupt 

Day  of  fast 

N 

P2O5 

Day  of  fast 

N 

P2O5 

1 

13.545 

2.597 

1 

10.01 

1.56 

2 

12.586 

2.925 

2 

9.92 

1.89 

3 

13.121 

3.289 

3 

13.29 

2.53 

4 

12.393 

2.974 

4 

12.78 

2.36 

5 

10.695 

2.871 

5 

10.95 

2.19 

6 

10.100 

2.667 

6 

9.88 

2.29 

7 

10.885 

2.663 

8 

8.903 

1.722 

9 

10.833 

2.065 

10 

9.467 

0.948 

The  average  daily  fecal  nitrogen  and  phosphorus  (P2Og)  with 
Cetti  were  0.316  gm.  and  0.205  gm.,  respectively,  and  with  Breit- 
haupt, 0.113  gm.  and  0.140  gm.,  respectively. 

The  phosphorus  of  the  urine  was  found  to  be  largely  combined 
with  potassium,  more  than  twice  as  much  as  was  present  in  com- 
bination with  calcium  and  magnesium.  Calcium  and  magnesium 
were  present  in  the  urine  only  as  phosphates.  By  computation 
the  authors  show  that  it  would  have  been  impossible  for  the  flesh  to 
have  given  up  all  of  the  phosphorus  excreted.  In  both  cases  a  part 
of  the  phosphorus  is  shown  to  have  come  from  the  bones. 


440 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


Gusmitta  (1893)  studied  the  composition  of  the  bones  of  sev- 
eral rabbits  and  a  dog  as  affected  by  starvation.  The  method  was 
amputation  of  parts,  and  comparison  with  others  remaining  after 
recovery  of  the  animal  from  the  mutilation,  and  after  subsequent 
starvation  to  death. 

The  bones  were  shown  to  lose  during  starvation,  in  weight, 
volume  and  specific  gravity,  and  to  increase  in  porosity ;  there  was  a 
slight  increase  in  water,  and  a  uniform  decrease  in  all  organic  and 
inorganic  constituents.  The  results  seem  indisputable  in  showing 
that  starvation  causes  katabolism  of  bones  as  well  as  soft  parts. 

The  following  figures  set  forth  results  with  the  dog  experi- 
mented upon. 
CONSTITUENTS  OF  BONES  OF  A  DOG  AS  AFFECTED  BY  STARVATION 


Dry  weight  of 
bones 

Total 

inorganic 

matter 

Iron 
phosphate 

Calcium 
carbonate 

Magnesium 
phosphate 

Calcium 

phosphate 

Grams  Percent 

Grams 

Percent 

Grams 

Percent 

Grams 

Percent 

Grams 

Percent 

Rig-ht  cubitus   and 
radius,  dog-  in  nor- 
mal state,  11.917 
g-m. 

Left    cubitus    and 
radius,  after  star- 
vation, 10.382  gm. 

7.261 
6.556 

60.93 
63.16 

0.1090* 
0.0134 

0.16 
0.13 

0.6888 
0.5391 

5.78 
5.20 

0.1120 
0.0809 

0.94 
0.78 

6.4208 
5.9011 

53.88 
56.84 

*  Apparently   erroneous;    0.0169  would  harmonize  with  percentage   figure. 

I.  Munk  (1894)  reported  a  fasting  experiment  with  a  dog 
which  shows  more  clearly  still  than  his  previous  experiment  on  Cetti 
that  in  inanition  a  part  of  the  nitrogen,  calcium,  magnesium  and 
phosphorus  loss  is  from  the  bones.  Some  of  his  figures  are  as  fol- 
lows: 

EXCRETION  OF  NITROGEN  AND  PHOSPHORUS  BY  A  MATURE 
FASTING  DOG— Grams 


Body  weight 

Day 

Kg. 

N 

P2O5 

P205:N 

Prelim. 

17.16 

16.82 

1.99 

1:8.4 

1 

16.94 

5.59 

0.820 

1:6.8 

2 

16.48 

5.31 

0.901 

1:5.9 

3 

15.85 

5.32 

0.996 

1:5.3 

4 

15.58 

5.28 

1.106 

1:4.8 

5 

15.35 

5.18 

1.268 

1:4.1 

6 

15.16 

4.29 

0.776 

1:5.5 

7 

14.95 

4.66 

1.020 

1:4.5 

8 

14.77 

3.79 

1.210 

1:3.1 

9 

14.49 

3.59 

0.883 

1:4.1 

10 

14.27 

3.74 

0.851 

1:4.4 

Total  loss 

2.89 

46.75 

9.83 

1:4.8 

Average 

0.29 

4.68 

0.98 

Feces 


Total 
Average 


1.576 
0.158 


1.874 
0.187 


1:0.84 


PHOSPHORUS  METABOLISM 


441 


During  the  preliminary  period  of  several  days  the  diet  was 
liberal  and  normal;  ratio  of  P205:N  in  urine  as  1:8.4.  During  fast 
this  ratio  was  1 :4.8.  The  increased  phosphorus  in  the  urine,  the 
author  computes,  would  be  equal  to  that  in  32  pi.  fresh  bone ;  the 
extra  calcium  would  imply  the  destruction  of  31.5  gm.  fresh  bone, 
and  an  excess  of  magnesium  was  also  to  be  accounted  for. 

Heymans's  (1896)  work  on  metabolism  during  starvation  of  the 
rabbit  shows  that  there  is  a  premortal  rise  in  phosphorus  excretion. 
If  the  fast  is  discontinued  by  the  giving  of  food  the  phosphorus 
excretion  in  the  urine  is  very  much  less  than  either  the  normal  or 
the  fasting  excretion,  until  an  equilibrium  is  reestablished. 

Weiske  (1897),  experimenting  with  rabbits,  studied  the  effects 
of  starvation  on  the  composition  of  the  bones,  teeth  and  other 
tissues.  Four  rabbits  from  the  same  litter  were  used,  No.  1  as  a 
control,  being  killed  at  the  beginning  of  the  experiment,  and  the 
three  others  after  fasting  for  7-11  days.  Below  are  some  of  the 
data  from  the  analysis  of  the  carcasses. 

COMPOSITION  OF  BODIES  OF  RABBITS  AS  AFFECTED  BY 
STARVATION 


No.  1 


No.  2 


No.  3 


No.  4 


Loss  of  Different  Parts — Percent 


Flesh 

Skeleton. . . 

Pelt 

Stomach. . . 
Intestines , 

Liver 

Lungs 

Kidneys. . . 

Heart 

Spleen 


39.7 

45.0 

4.1 

Gain 

23.3 

20.3 

20.8 

34.8 

24.7 

50.4 

56.5 

54.4 

68.1 

62.5 

26.3 

29.5 

29.7 

23.4 

73.9 

69.6 

43.4 
3.3 
20.0 
27.7 
48.2 
62.5 
53.3 
26.9 
Gain 
71.7 


Weight  of  Parts  in  Grams 


Flesh,  water  and  fat-free 

Skeleton,  water  and  fat-free. 

Fat  in  flesh 

Fat  in  skeleton 

Fat  in  flesh  and  skeleton 


210.33 

147.00 

133.39 

88.79 

95.65 

101.26 

41.32 

4.75 

4.98 

11.58 

0.52 

0.53 

52.90 

5.27 

5.51 

Analysis  of  Bones  Other  Than  Leg  Bones — Percent  of  Water-  and  Pat-free 


Organic  substance  , 
Mineral  substance. . 

CaO 

MgO 

COS 

P205 

Remainder 


38.06 

39.73 

37.74 

61.94 

60.27 

62.26 

32.35 

31.76 

32.52 

0.71 

0.69 

0.62 

2.50 

2.70 

2.88 

24.17 

24.02 

24.67 

2.21 

1.10 

1.57 

39.35 
60.65 
31.77 
0.64 
2.48 
24.22 
1.54 


Analysis  of  Teeth — Percent  of  Water-  and  Pat-free 


Organic  substance. 
Mineral  substance. . 

CaO 

MgO..„ 

CO2 

P2O5 

Remainder 


23.42 

21.80 

21.52 

76.58 

78.20 

78.48 

37.88 

38.75 

38.92   - 

2.49 

2.44 

2.49 

0.77 

0.99 

1.10 

33.91 

34.21 

34.46 

1.53 

1.81 

1.52 

22.09 

77.91 

38.83 

2.37 

1.45 

34.24 

1.02 


442 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BTJL.  5 


It  will  be  noted  that  the  leg  bones  were  not  included  in  the  above 
analyses.  In  an  effort  to  harmonize  these  results,  which  do  not 
show  a  loss  of  phosphorus  from  the  bones,  with  those  which  have 
been  discussed,  we  can  only  suggest  that  the  bones  constitute,  rela- 
tively, so  large  a  store  of  the  mineral  constituents  of  the  body  that  a 
considerable  change  in  amounts  of  urinary  constituents  would  repre- 
sent but  a  slight  change  in  the  skeleton  as  a  whole,  so  slight  a 
change,  in  fact,  that  its  detection  by  skeleton  analysis  would  be  very 
likely  to  be  missed  through  the  inevitable  errors  involved  in  the 
comparison  of  a  starved  animal  with  another  individual  serving  as  a 
control ;  further,  the  fasting  time  was  short ;  the  skeleton  yields  but 
slowly  to  influences  which  tend  to  modify  its  composition. 

Hoover  and  Sollmann  (1897)  studied  phosphorus  metabolism 
during  fasting  in  hypnotic  sleep  covering  a  period  of  8  days.  The 
analysis  of  the  urine  is  as  follows : 

ANALYSIS  OF  URINE  DURING  HYPNOTIC  FAST— Grams 


Date 

Quantity 
C.C. 

Urea 

Uric  acid 

P2O5 

NaCl 

Total  N 

Water  taken 

July  15 

1350 

36.21 

0.824 

3.381 

12.330 

20.978 

.J 

"    16 

570 

22.62 

0.617 

2.303 

6.837 

12.369 

..     1? 

470 

22.99 

0.450 

2.268 

3.863 

12.370 

750 

"    18 

530 

25.24 

0.538 

2.270 

3.964 

14.013 

750 

"19  &  20 

1020 

58.18 

0.866 

5.052 

5.954 

27.988 

1150 

"    21 

410 

20.67 

0.375 

2.434 

2.496 

10.791 

875 

"    22 

660 

28.26 

0.572 

3.150 

2.419 

14.504 

1250 

"    23 

820 

34.13 

0.908 

4.442 

3.296 

21.582 

750 

The  loss  in  weight  during  the  last  7  days  was  5896  gm. ;  loss  of 
nitrogen,  113.617  gm. ;  nitrogen  estimated  as  albumin  710  gm., 
— estimated  as  muscle  3341  gm. 

There  was  some  evidence  of  an  increased  proportion  of  phos- 
phorus to  nitrogen  during  the  progress  of  the  fast.  The  average 
loss  of  phosphorus  during  the  last  7  days  was  3.131  gm.  P205. 

Nemser  (1899)  studied  the  effects  of  starvation  on  the  total  and 
nuclein  phosphorus  content  of  the  muscles,  liver,  kidney  and  intes- 
tine of  white  mice.  Eighty  mice  were  used.  The  data  seem  to 
indicate  that  the  percent  of  loss  in  weight  of  parts  was  greater  than 
the  gain  in  percent  of  total  and  nuclein  phosphorus,  which  implies 
that  the  total  and  nuclein  phosphorus  contributed  to  the  loss  during 
fast. 

Sedlmair  (1899)  studied  the  losses  sustained  by  the  various 
organs  of  the  cat  during  starvation,  by  comparing  three  cats  of  the 
same  litter,  one  well-fed,  with  two  which  were  starved.  One  cat, 
starved  for  36  days,  and  losing  in  weight  1764  gm.  from  an  initial 


PHOSPHORUS  METABOLISM 


443 


weight  of  3368  gm.,  apparently  lost  in  this  time  only  0.564  gm.  CaO, 
which  was  computed  to  represent  3.4  gm.  of  fresh  bone  substance, 
or  less  than  1  percent  of  the  total  weight  of  the  skeleton.  Below 
are  figures  which  distribute  the  total  loss  to  the  various  parts  sus- 
taining decrease  in  weight. 

LOSSES  IN  DRY  SUBSTANCE  OF  DIFFERENT  TISSUES  OF  CATS 
DURING  STARVATION— Percents  of  Total  Loss 


Cat  B 


Cat  C 


Muscular  system 

Skin 

Mesentery 

Bones ■ 

Liver 

Intestines 

Kidneys 

Pancreas 

Heart 

Lungs 

Spleen 

Brain 

Spinal  marrow 

Central  nervous  system 

Urinary  bladder,  aorta,  trachea  and  eyes 
Blood 


62.23 

57.06 

11.33 

16.50 

8.51 

7.56 

5.67 

8.17 

5.03 

4.41 

3.00 

3.18 

0.80 

0.73 

0.22 

0.41 

0.39 

0.31 

0.24 

0.30 

0.25 

0.25 

+0.17 

+0.10 

0.05 

0.07 

+0.12 

+0.03 

0.16 

0.06 

2.29 

1.11 

100.00 


100.02 


The  bones  were  in  most  cases  made  richer  in  water  by  starva- 
tion, while  the  dry  substance  decreased,  both  absolutely  and  in  per- 
cent, these  effects  being  most  noticeable  in  the  long  bones.  The 
principal  loss  of  weight  from  the  bones  was  fat,  though  all  other 
constituents  of  the  bones  shared  in  the  loss. 

Schulz  and  Mainzer  (1901)  studied  nitrogen  and  phosphorus 
metabolism  in  3  starving  rabbits  and  1  starving  dog.  They  observed 
in  each  case  the  usual  premortal  rise  in  the  urinary  excretion  of 
both  nitrogen  and  phosphorus,  but  no  significant  change  in  ratio  of 
nitrogen  to  phosphorus  in  the  urine  just  prior  to  death.  Death 
occurred  before  the  disappearance  of  fat  from  the  body. 

Rubow  (1905)  found  the  lecithin  content  of  9  normal  dogs' 
hearts  to  vary  between  7.05  and  8.80  percent ;  while  of  fat  the  same 
hearts  contained  3.60  to  3.99  percent,  in  the  dry  substance.  Two 
dogs  which  were  starved  to  death,  had  in  the  hearts  7.3  and  7.67 
percent  of  lecithin,  and  the  same  hearts  contained  3.67 — 3.71  percent 
of  fat,  on  the  dry  basis.  In  normal  dog  muscle  the  lecithin  varied 
between  4.44  and  5.27  percent,  while  the  fat  content  of  the  same 
samples  was  14.16  and  6.30  percent,  respectively,  on  the  dry  basis. 
In  the  muscle  of  the  starved  dogs  was  found  3.08  and  3.74  percent  of 
lecithin,  and  2.58  and  2.74  percent  of  fat,  on  the  dry  basis. 

From  these  figures  it  would  appear  that  in  starvation  the  dog 
uses  both  fat  and  lecithin  from  the  muscles,  but  the  withdrawal  of 
either  fat  or  lecithin  from  the  heart  is  doubtful. 


444 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


Van  Hoogenhuyze  and  Verploegh  (1905),  in  studying  creatinin 
excretion,  publish  urine  analyses  of  a  human  subject,  covering  15 
days  of  fast,  preceded  and  followed  by  one  day  with  food.  The 
figures  are  as  follows: 

URINE  ANALYSIS  OF  A  FASTING  MAN— Grams 


Date 

Total  N 

Creatinin 

P2O5 

X/P2O5 

Remarks 

June  10 

13.99 

1.087 

2.670 

5.2 

Food 

11 

8.76 

0.904 

1.550 

5.8 

Beginning-  of  starvation  period 

"    12 

8.38 

0.577 

1.830 

4.6 

'    13 

10.73 

0.581 

2.654 

4.0 

'    14 

9.40 

0.634 

2.934 

3.2 

In  the  morning-,  50  c.c.  sulphatic  water  drunk 

'    15 

7.87 

0.603 

1.749 

4.5 

In  the  morning-,  50  c.c.  sulphatic  water  drunk 

'    16 

7.73 

0.590 

1.069 

7.2 

In  the  morning-,100  c.c.  sulphatic  water  drunk 

'    17 

6.11 

0.469 

0.713 

8.5 

From  11  to  1  o'clock,  muscular  work 

'    18 

7.70 

0.689 

1.658 

4.6 

19 

7.35 

0.715 

1.702 

4.3 

20 

6.80 

0.602 

1.461 

4.6 

'    21 

6.14 

0.453 

1.097 

5.6 

'    22 

6.97 

0.566 

1.312 

5.3 

'    23 

5.62     . 

0.548 

1.114 

5.0 

'    24 

4.08 

0.426 

0.869 

4.7 

'    25 

4.38 

0.715 

0.539 

8.0 

Food  taken  at  10  o'clock  at  night 

'"    26 

7.23 

1.028 

0.145 

49.9 

Food 

At  the  beginning  of  this  fast  the  nitrogen,  phosphorus  and 
creatinin  decreased  from  the  quantities  excreted  during  the  day 
before,  and,  as  tissue  protein  was  attacked,  the  ratio  of  N  to  P205 
in  the  urine  became  less.  The  muscular  work  of  the  7th  day 
of  fasting  had  the  effect  temporarily  to  lessen  the  excretion  of  nitro- 
gen, creatinin  and  phosphorus,  especially  phosphorus,  though  the 
excretion  of  all  these  constituents  rose  on  the  next  day,  the  creatinin 
and  phosphorus  elimination  remaining  high  for  three  days.  At 
the  end  of  the  fast  the  total  nitrogen  and  creatinin  excretions  rose 
markedly,  but  the  phosphorus  excretion  sank  to  a  lower  figure  than 
at  any  time  during  the  fast,  or  on  the  day  before  the  fast,  which 
implies  a  lessened  katabolism  of  phosphorized  proteins,  and  the 
replenishment  of  depleted  phosphorus  reserves. 

Cathcart  and  Fawsitt  (1907)  reported  on  a  14-day  fasting 
experiment,  with  a  31-year-old  man,  with  a  3-day  preliminary  and 
5-day  after-period,  in  which  nearly  one-fifth  of  the  phosphorus 
seemed  to  have  some  source  other  than  the  soft  parts.  During  this 
fast  the  elimination  of  purin  bases  and  phosphorus  decreased  on  the 
first  day,  the  organism  apparently  sparing  its  nucleoproteins.  The 
minimum  purin  excretion  was  on  the  third  day  of  the  fast,  after 
which  it  increased,  reaching  the  normal  on  the  tenth  day.  The 
phosphorus  outgo,  however,  steadily  decreased,  as  also  did  total 
nitrogen,  and  both  were  less  in  amount  on  the  last  day  of  the  fast 
than  on  any  other.  Thus  purins  increased  as  phosphates  and  total 
nitrogen  decreased.  That  nucleins  participated  in  the  tissue 
katabolism  is,  of  course,  clear.  The  fate  of  the  nonpurin  nitrogen 
and  of  the  phosphorus  from  the  nucleins,  however,  is  not  so  certain, 


PHOSPHORUS  METABOLISM  445 

but  since  the  outgo  of  total  nitrogen  and  phosphorus  was  under- 
going a  progressive  decrease  it  would  be  fair  to  assume  that  these 
constituents  were  retained,  or  else,  if  eliminated,  that  they  served 
to  protect  equivalent  amounts  of  the  same  in  a  higher  state  of 
organization  or  combination. 

Wellman  (1908)  studied  mineral  metabolism  in  fasting  rabbits. 
In  experiments  of  12-15  days  duration  it  was  determined  that  full- 
grown  rabbits,  in  starving  to  death,  lost  6.5  to  7.7  percent  of  the 
fat-free,  dry  substance  in  the  bones,  and  about  14  percent  of  the 
fresh  weight.  Of  this  loss  of  fresh  weight  nearly  half  was  fat. 
The  absolute  loss  of  calcium  from  the  bones  was  determined  as  1.8 
gm.,  of  phosphorus  0.6  gm.,  and  of  dry  substance  8.7  gm. 

Falta  and  Whitney  (1908)  also  found  in  the  fasting  metabolism 
of  the  dog  such  an  increase  in  the  proportions  of  Ca  and  P  to  N  in 
the  urine  as  indicated  a  probable  participation  of  the  bones  in  the 
increased  katabolism. 

Grund  (1910)  concluded  that  the  ratio  P:N  in  the  organs  of 
animals  tends  strongly  to  remain  constant  under  different  conditions 
of  nourishment  as  well  as  in  fasting,  though  some  variation  in  this 
proportion  was  noted  in  the  liver  of  the  hen  and  the  dog.  Accord- 
ing to  recent  observations  (1912a,  1912b,  1913)  the  relative  amount 
of  protein  phosphorus  in  muscle,  in  relation  to  total  phosphorus  and 
to  the  protein  nitrogen,  is  not  altered  by  the  wasting  away  due  to 
hunger,  but  is  materially  increased  by  degeneration  due  to  the  sever- 
ing of  nerves  or  to  severing  from  the  bone.  Atrophy,  from  division 
of  the  nerve,  and  from  severing  from  the  bone,  alike,  cause  a  dissolv- 
ing of  phosphorus-free  proteins,  with  a  sparing  of  phosphoproteins. 

Wolf  and  (Esterberg  (1911)  made  a  study  of  nitrogen,  sulphur 
and  phosphorus  metabolism  in  the  dog  as  affected  by  fasting  or 
underfeeding.  The  feeding  of  a  comparatively  small  amount  of  pro- 
tein was  found  to  reduce  the  phosphorus  excretion  to  a  very 
low  amount.  The  feeding  of  starch  and  fat  seemed  to  have  little  or 
no  effect  on  phosphorus  excretion.  During  fasting  the  ratio  of 
phosphorus  to  nitrogen  in  the  urine  was  so  high  that  it  seemed  prob- 
able that  some  of  the  phosphorus  came  from  the  bones. 

Wilson  and  Hawk  (1914)  found  that  during  fast  the  urinary 
acidity  and  phosphates  varied,  in  general,  together,  increasing 
amounts  of  acid  in  the  blood  or  lymph  (due  to  increased  fat 
katabolism  in  the  presence  of  decreased  carbohydrate  oxidation) 
leading  to  a  change  of  di-  to  monophosphates,  and  the  excretion  of 
the  excess  of  the  latter.  Thus  increased  phosphate  excretion  and 
urinary  acidity  followed  an  increased  formation  of  acids  in  the 
body.  Increased  neutralization  of  acids  with  ammonia  led  to 
decreased  urinary  acidity  and  phosphates. 


446  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Among  the  other  changes  produced  by  starvation,  in  the  phos- 
phorus compounds  of  animals,  may  be  mentioned  the  reduction  in 
the  lecithin  phosphorus  of  the  liver  as  noted  by  Heffter  (1890, 1891) 
and  the  decrease  in  the  phosphocarnic  acid  in  the  brain  as  observed 
by  Panella  (1906a). 

Summary.  Fasting  causes  an  increased  outgo  of  phosphorus. 
From  the  evidence  that  non-purin  nitrogen,  as  well  as  nuclein  com- 
pounds and  skeleton,  all  contribute  to  this  outgo  we  conclude  that 
its  production  is  due  to  generalized  katabolism.  The  proportionate 
losses  sustained  by  the  various  tissues  are  difficult  of  determination. 
The  loss  from  the  skeleton,  however,  is  appreciable,  though  but  a 
very  small  part  of  the  total  phosphorus  present.  The  lecithin  phos- 
phorus of  the  liver  and  muscles,  and  the  phosphocarnic  acid  of  the 
brain  have  been  shown  to  contribute  to  the  phosphorus  loss  in  star- 
vation. 

There  is  evidence  that  the  first  deficiencies  of  phosphorus  are 
met  principally  by  non-purin  compounds,  that  there  is  a  progressive 
decrease  of  phosphorus  outgo  from  this  source,  and  also  of  total 
phosphorus,  until  the  premortal  rise,  but  increasing  contributions 
from  nuclear  and  skeletal  compounds. 

There  is  evidence  of  a  further  utilization  by  the  animal  of  the 
phosphorus  of  katabolized  nuclein  compounds  after  the  elimination 
of  the  purin  nitrogen. 

The  feeding  of  protein  during  starvation  has  a  much  greater 
effect  to  reduce  phosphorus  loss  than  the  feeding  of  starch  and  fat. 

PHOSPHORUS  METABOLISM  DURING  INCUBATION 

As  typical  of  metabolism  generally,  especial  interest  attaches 
to  the  simplified  physiology  of  the  egg.  C.  Voit  (1877a)  sought  to 
determine  if  there  is  a  loss  of  lime  from  the  egg-shell,  and  utiliza- 
tion of  the  same  in  the  growth  of  the  skeleton  of  the  incubating 
chick.  He  concluded  that  there  was  no  such  loss  of  lime  from  the 
shell,  but  his  method  of  demonstration  leaves  one  in  doubt  as  to  the 
fact. 

During  incubation  A.  Kossel  (1885)  found  that  the  phospho- 
protein  of  the  hen's  egg  changes  to  nucleoprotein,  because,  while  the 
yolk  nuclein  of  the  fresh  egg  does  not  yield  purin  bases,  the  nuclein 
of  the  hatched  chick  does  yield  guanin  and  hypoxanthin. 

Maxwell  (1893)  studied  the  phosphorus  exchanges  in  the  incu- 
bating hen's  egg  but  the  analytical  data  indicate  that  his  methods 
were  unsatisfactory. 


PHOSPHORUS  METABOLISM 


447 


Mesernitzy  (1907)  states  that  the  lecithin  of  the  fresh  hen's 
egg,  dried,  is  about  15.35  percent,  estimated  from  the  phosphorus  of 
the  ether  extract.  During  incubation  he  found  this  amount 
reduced  by  about  a  half. 

Carpiaux  (1908)  shows  that  with  the  decrease  of  lecithin  in  the 
incubating  hen's  egg  there  proceeds  an  accompanying  decrease  in 
the  lime  of  the  shell,  and  an  increase  of  inorganic  phosphates.  The 
following  data  substantiate  these  conclusions. 

ANALYSES  OF  HENS'  EGGS  DURING  INCUBATION— Grams 


Exp.  No. 

Agre  in 

Weight 

CaO 

P2O5, 

P2O5, 

P2O5, 

Lecithin 

days 

inorganic 

organic 

total 

1 

0 

0.040 

0.075 

0.150 

0.225 

1.050 

2 

6-7 

62.63 

0.128 

0.146 

0.274 

1.022 

3 

6-7 

55.43 

0.0377 

. 

4 

7-8 

64.70 

0.0360 

0.127 

0.135 

0.262 

0.945 

5 

13-14 

54.20 

0.0478 

6 

14-15 

54.62 

0.0616 

0.137 

0.108 

0.245 

0.760 

7 

18 

55.90 

0.1357 

0.182 

0.050 

0.232 

0.350 

8 

19 

53.10 

0.1487 

9 

21 

0.2022 

Plimmer  and  Scott  (1909)  compared  the  phosphorus  distribu- 
tion in  the  body  of  the  chick  at  the  time  of  hatching  with  that  of 
the  fresh  egg,  and  of  the  egg  at  different  times  during  incubation. 
They  draw  the  following  conclusions  with  regard  to  the  sources  of 
the  different  phosphorus  compounds  of  the  young  bird's  body : 

"There  is  not  much  change  in  the  ether-soluble  phosphorus 
until  about  the  16th  or  17th  day  of  incubation,  when  a  very  rapid 
disappearance  of  the  ether-soluble  phosphorus  bodies  commences. 
At  this  time  it  corresponds  with  a  great  increase  in  the  amount  of 
inorganic  phosphate  in  the  chicken;  therefore  it  is  impossible  to 
avoid  the  conclusion  that  these  glycerophosphoric  acid  compounds 
have  been  converted  into  inorganic  phosphate  for  the  calcification 
of  the  bones ....  Some  of  the  inorganic  phosphate  must  have  come 
from  the  vitellin,  since  all  of  it  disappeared.  Probably  some  of  it, 
however,  was  converted  into  nucleic  acid,  as  the  latter  increased .... 
There  is  a  gradual  absorption  of  the  protein  phosphorus  bodies  of 
the  yolk  by  the  developing  chicken  before  there  is  a  change  in  the 
lecithin  bodies." 

General  Conclusion:  "That  the  glycerophosphoric  acid  gives 
rise  only  to  inorganic  phosphate  in  the  developing  chicken  and  is  not 
transformed  into  any  combination  with  protein.  There  is  no  evi- 
dence of  a  synthetic  process  occurring  in  the  developing  egg  as 
regards  the  phosphorus  compounds  unless  the  probable  transforma- 
tion of  the  phosphoprotein  into  nucleoprotein  be  so  considered." 

Data  from  this  study  are  in  the  following  table. 


448 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


FH 

£ 

bo 

w 

bo 

M 

W 

W 

cw 

© 

In 

hH 

n 

0) 

H 

«* 

85 

C/2 

O 

^S 

o 

u 

o 

w 

H 

«! 

ffl 

bo 
bo 
W 

S 

i— i 

U 

i— i 

•  F* 

CO   ft: 

CO 

PO 

S-H 

« 

rp 

Oh 

o 
W 

Q 

n 

CO 

3 

cu 

i— i 

S-i 

CO  03 

iTi 

O  W 

O. 

W  Oh 

to 

n 

Oh 

fc 

O 

S3 

£ 

H 

o 

«M 

H 

P 

co 

M 

s 

HH 

CU 

(3 

w 

H 

<u 

CO 

£U 

a 
"3 
> 

°bo 

.    .    .10    .cocm   .    . 

.     •     -00     -lOCO     •     • 
•      ■      -iH      T-lr-(      •      • 

o 

■     •     -O      -OO     ;0 

ho 

ho 

H(DO     .      .     ..-1     .      . 

t^COCO      •      •     •  i— <     •      • 
CMCNCM     •      •     "rH     ■      • 

1-° 

o  2 

.9-- 

<0.S 

_d  <u 

'3  "o 

S  * 

en  '-3 

33 

"3  .. 

jj  ho 
to  ho 

.     .     .  •*     .00£>.     .     . 

.     •     ."*31     <  010     •      • 
•     •     ■  CM     "CMCM     •     • 

o 

o 

.     .      .10     .<MO     .O 

.    .    .^    .r^oo    -cm 

•     •      -CM      •CM'-H     -rH 

ho 
ho 

O^HO     .10      .OOi— 1    .        ' 

a 
"o 

0) 

u 

°ho 

in  ho 

O   0) 

•      .     .'tH     -OOO     ■     • 
.      .      -co     -50CO     •      - 

3 
o 

.    .    .m    .coco    -co 

•     •      -CM     -rHi-1     'iH 

ho 
ho 

s 

OOi— IO      .CD     .OO     • 

•*-*CO      -CN)     '"-ICO     • 

cococo    *co    *ioco    ■ 

.3 

i 

J3  5 
CV+J 

S3 

■d  o 
p,») 

o*d 

'3 '3 

ho 
ti 
o 

a 

►H 

v  ho 
en  ho 

.    .     .  o    .  0  o    .    . 
'■'■'.  u    '■  t~  u    '■    • 

o 

3 
o 

.     .     .CM     .IOCO     .O 

•  •     'CO     -OO      'O 

•  •     •!-!     -COIO     -CO 

ho 
ho 

Trace 

Trace 

Present 

Present 

19!  i 

47.7 

d 

_  to 

o  o 
Eh  « 

d 

4j  ho 
tf>  ho 
u  cu 

.      .      .00      .Tf(^H     .      . 

•    •    -co    "o  ^    •    • 

3 
o 

.      .     .O     .OtJI     .CO 

•    .    -co    -inr^    'Go 

»      •     'iO     -IOCD     *CD 

ho 
ho 

(MCM"— 1      -CM     .(MO)     • 

coooo>    'O    •r^o)    • 
rH     -i-H     -OJlO     • 

t 

d 
S.2 

After  7  days 

"    13    "     

"    14    "     

"    16    '"     

"    17    ;      

"   19    ;   

"    20           

PHOSPHORUS  METABOLISM 


449 


Hanes  (1912a,  1912b)  finds,  by  microchemical  studies,  that 
phosphorized  fats  are  abundant  in  the  liver  of  the  incubating  chick 
during  the  first  two  weeks ;  during  the  third  week  these  compounds 
diminish,  the  phosphorus  apparently  serving  for  calcification.  It 
is  suggested  that  the  pathological  calcification  of  arteriosclerosis 
results  from  a  splitting  in  situ  of  phosphorized  fats,  with  subsequent 
formation  of  calcium  salts.      See  also  Tornani  (1909). 

Robert  and  Wasteneys  (1913)  concluded  that  serious  errors 
vitiated  the  results  of  Massing  and  of  Shackell  in  their  studies  on 
the  phosphorus  changes  in  developing  sea-urchin  eggs.  Frdm  an 
investigation  of  their  own  they  submit  the  following  data : 

FORMS  OF  PHOSPHORUS  IN  DEVELOPING  SEA-URCHIN  EGGS 
Percent  of  Total  P 


Stage  of  development 

Soluble  in  alcohol 

Soluble  in  water 

Insoluble 

Exp.  1 

Exp.  2 

Exp.  1 

Exp.  2 

Exp.  1 

Exp.  2 

39.5 
36.5 
35.2 

46.5 
38.8 
35.1 

36.7 
47.6 
30.8 

29.6 
40.6 
37.8 

23.8 
15.9 
34.0 

23.9 

20.6 

Plutei 

27.1 

The  conclusions  are  as  follows: 

"1.  During  the  development  of  the  eggs  of  Strongylocentrotus 
purpuratus  to  blastulae  the  proportion  of  P  which  is  present  in  the 
form  of  phospholipines  (lecithin,  etc.)  undergoes  appreciable  dimin- 
ution. The  further  development  of  the  blastulae  into  plutei  is 
accompanied  by  a  further  diminution  in  the  proportion  of  phospho- 
lipines in  the  larvae. 

"2.  The  development  of  the  eggs  of  S.  p.  to  blastulae  is  accom- 
panied by  a  considerable  increase  in  the  proportion  of  P  which  is 
soluble  in  boiling  water  and  insoluble  in  alcohol,  and  by  a  decrease 
in  the  proportion  of  P  which  is  insoluble  either  in  alcohol  or  in  boil- 
ing water.  The  further  development  of  the  larvae  to  plutei  is 
accompanied,  on  the  contrary,  by  a  decrease  in  the  proportion  of 
water-soluble  phosphorus  and  an  increase  in  the  proportion  of  insol- 
uble P." 


PHOSPHORUS  METABOLISM  IN  INFANCY 

RELATIVE  VALUE  OF  DIFFERENT  KINDS  OF  MILK 

Human  milk  is  low  in  the  ash  constituents  generally,  and  also 
low  in  phosphorus,  as  compared  with  all  other  milks.  It  is  also  low 
in  casein  as  compared  with  most  of  them,  including  cow's  milk.    It 


450  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

has,  however,  a  relatively  high  content  of  both  phosphatid  and 
nucleon,  and  of  Wroblewski's  opalisin,  and  a  relatively  large  part  of 
its  phosphorus  is  in  an  organic  form.  Each  of  these  items  has  been 
brought  forward  as  an  explanation  of  the  undoubted  fact  that  child- 
ren thrive  better,  and  form  bones  and  tissues  more  rapidly,  when 
they  are  fed  on  mother's  milk  than  when  artificially  fed. 

In  composition,  the  milk  of  equidae  is  most  nearly  like  human 
milk.  Schlossmann  (1897)  considered  ass's  milk  with  a  view  to  its 
value  as  a  substitute  for  human  milk,  but  decided  that  it  would  be 
unsatisfactory  on  account  of  its  high  proportion  of  protein  to  fat. 

Aside  from  differences  in  amounts,  the  caseins  are  found  to  be 
different  in  kind,  the  evidences  being  reported  in  our  discussion  of 
the  chemistry  of  casein  on  pages  43  and  44. 

Soxhlet  (1893)  discusses  differences  between  human  and  cow's 
milk,  with  consideration  of  suitable  modifications  of  cow's  milk  to 
prepare  it  for  the  human  infant.  He  says  that  cow's  milk  gives 
coarser  clots  with  rennet  ferment,  and  contains  about  twice  as  much 
casein  and  6  times  as  much  calcium,  and  has  3  times  as  high  acidity, 
as  human  milk.  Addition  of  water,  or  heating  the  cow's  milk, 
reduces  the  size  of  colts ;  but  in  each  case  other  effects  produced  are 
unfavorable.  Decoctions  of  grains,  etc.,  affect  the  clots  as  do  corre- 
sponding amounts  of  water,  but  add  starch ;  if  malt  extract  be  added 
the  resultant  mixture  may  be  of  higher  nutritive  value.  Since  cow's 
milk  contains  4  times  as  much  phosphoric  acid  and  6  times  as  much 
calcium  as  human  milk,  more  calcium  phosphate  is  supplied  than  is 
needed  for  bone-formation,  and  the  excess  is  excreted,  mostly  as 
calcium  soaps  in  the  feces.  Both  kinds  of  milk  contain  the  essen- 
tial soluble  calcium,  principally  in  the  form  of  calcium  citrate. 

Julius  Lehmann  (as  reported  by  Hempel,  1894)  and  Hammar- 
sten  (1895)  both  proposed  modification  of  cow's  milk  for  the  use  of 
human  infants,  on  the  basis  of  the  consideration  that  the  most 
significant  difference  between  human  and  cow's  milk  is  in  the  ratio 
of  albumin  to  casein,  and  of  casein  to  fat. 

Stoklasa  (1897)  gives  the  range  of  lecithin  in  cow's  milk  as 
0.90-1.13  gm.  per  liter  (representing  an  average  of  0.091  gm.  P205), 
and  that  in  human  milk  as  1.70-1.86  gm.  per  liter  (representing  an 
average  of  0.153  gm.  P205).  In  cow's  milk  the  total  P205  is  1.81  gm. 
per  liter,  and  therefore  the  lecithin  phosphorus  forms  5  percent  of 
the  total ;  while  in  human  milk  there  is  but  0.44  gm.  P205  per  liter, 
and  the  lecithin,  therefore,  forms  a  much  larger  proportion,  35  per- 
cent, of  the  total. 


PHOSPHOKUS  METABOLISM 


451 


Blauberg  (1897c),  studying  the  nutrition  of  infants  on  woman's 
milk  and  on  cow's  milk  diet,  found  about  the  same  differences  in  the 
mineral  constituents  of  the  feces  as  in  ash  of  the  two  kinds  of  milk, 
especially  a  larger  calcium  and  phosphorus,  and  lower  iron  content 
in  the  feces  from  cow's  milk. 

Blauberg  (1897b)  reports  an  analysis  of  Mellin's  Food.  For 
comparison  with  these  figures  we  have  computed  to  the  dry  basis 
the  proximate  food  constituents  of  human  milk  as  reported  by 
Ellenberger,  Seeliger  and  Klimmer  (1902),  and  the  mineral  con- 
stituents as  reported  by  Soldner  (1902) .  From  these  data  it  would 
appear  that  the  use  of  Mellin's  Food  with  milk  would  tend  to 
substitute  carbohydrates  for  fats,  and  to  decrease  the  calcium.  The 
phosphorus  content,  on  the  dry  basis,  is  not  less  than  in  human 
milk,  and  as  Mellin's  Food  is  used  not  with  human  but  with  cow's 
milk,  which  contains  much  more  calcium  and  phosphorus  than  does 
human  milk,  it  seems  likely  that  the  use  of  this  proprietary  food 
would  not  have  the  effect  to  starve  the  infant  for  any  of  the  mineral 
nutrients. 


COMPARISON  OF  THE  DRY  MATTER  OF  MELLIN'S  FOOD  AND  HUMAN 

MILK 


Water, 
percent 

Percetnts  of  dry  substance 

Protein 

Fat 

Carbo- 
hy- 
drate 

Mineral 
matter 

K2O 

Na20 

CaO 

MgO 

P2O5 

CI 

Mellin's  food 
Human  milk 

6.31 

86.4 

8.2 
11.8 

2.22 
35.3 

85.15 

48.5 

3.93 

1.84 

1.15 
0.65 

0.41 
0.26 

0.025 
0.28 

0.047 
0.039 

.0.38 
0.23 

0.081 
0.43 

For  other  analyses  of  infant  foods  see  Blauberg  (1897a), 
Klautsch  (1896),  and  vonSzontagh  (1902). 

Keller  (1898)  determined  that  when  feeding  infants  on  cow's 
milk  more  phosphorus  is  found  in  the  urine  than  under  normal  feed- 
ing, and  also  a  very  much  larger  proportion  of  phosphorus  to  nitro- 
gen. 

Keller  (1900b)  studied  nitrogen  and  phosphorus  metabolism 
with  infants,  both  normal  and  suffering. from  gastrointestinal  dis- 
orders, on  normal  and  on  artificial  milk  feeding.  By  the  addition 
of  Na2H  P04  to  human  milk  the  phosphorus  retention  was  increased 
in  either  well  or  sick  infants.  Phosphorus  retention  was  greater  on 
human  milk  than  on  cow's  milk. 

Balance  data  from  Keller's  studies  are  in  the  following  table. 


'452 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


O 

S 

w. 

-3 

o 

M 
<! 
H 

H 

P 

O 

PC 

Pi 

CO 

o 

w 

Oh 

O 
J? 


I  5 


■J 


&  go 
O 


Q 
H 

CO 

<J   ■ 
"    i— i 


«3 


Q 


Q 

w 

H 

H  <! 
O  H 
•«  W 

5 


3 

One-third  cow's  milk  plus  Na2HP04 

Woman's  milk  plus  Na2HP04 

One-half  cow's  milk  plus  Na2HP04 

Woman's  milk 

Woman's  milk  plus  Na2HP04 

Woman's  milk  plus  Na2HP04 

Woman's  milk  plus  Na2HP04 

Woman's  milk 

Cow's  milk,  whole,  plus  Na2HP04 

Malted  milk  plus  Na2HP04 

Woman's  milk  plus  Na2HP04 

Two-fifths  cow's  milk 

One-third  cow's  milk  plus  Na2HP04 

o 

CO 

3 
u 

o 

43 

a 

to 
o 

XI 

A 

co  i   2- 

OIBfl 

OOC(3i-IWCClOW50lt50iCOr-.t^ 

OOCOCOr-COeOM-^OlOCOCOOO 
tHIOCOCOCOIQ-^TOCMi-ItJIt* 

o 

•S 

ioO'>*(Niecocoao-H'«i<coi-ieo 

mCSCOCOOQOCOCBlOINr- IrHCO 

nocflOiHocoocoinH'^n 

ooooooooooooo 

CO 
CO 

fa 

r»  <-h  co  o  o  cm  -h  -j  ai  t~  co  t~  o 

ooscoiomaocoirsos-^coooo 

HOCOOOOHOOOMOi-lO 

ooooooooooooo 

8 
fa 

05gt>.ogoccoinoQ'*iOi-[ 

OCOOOCO.-ICOaOCOOSOOCOOO''* 

ooooooooooooo 

0 

bo 
o 
+■> 

u  u  a 

fa     *> 

.-1  CO  Ifi  00  OS  OS  CO  r~  t»  CM  1(5  T*  00 

WaOOSmr^-HCO-HOi-H-WCOCT! 
i— ICO       miN>OCDlOH^l«Cl5H 

o 

.9 

P 

comcoo-^cooooooicooccDoo 

rHlOOt^OjMCOX3!»OCMm 

t-iC-cor-cDcoior-iooococooj 

<-IOi-IOOOOOCOOOrtO 

CO 
0) 

o 
S 

fa 

Tflomas^iflcOTWCooico.— ion 

CMO}t~Or-OCOTt<:j}OCO''*CO 
.— tCOC0CMr-HCOCM<MCM'*f«i— I.-C 

OOOOOOOOOOOOO 

fa 

OOCOOOOCOi-HOWiniOOOCOr- 
corHioocoocor^r^r^cMCMiO 

TflOOf'- ICOt^OOI—  OOmcOCO 

0? 

momoo 

CO  CO  COO  CO  00**  00  00      ■** 
t-ICOi— If— iCOCM^JICM-^CO.— iCOTti 

3690 
4190 
3960 
4380 
3190 
3300 
3530 
4350 
4380 
3640 
3630 
4900 
3550 

Agre  of 
child 

Mos. 

co  m  \a  co  cm  cm  cm  eo  <-i  10  ■*»  cm  ■<* 

Leng-th 
of  exp. 

Days 

WIOIOU51010  0U510IOI01010 

•a 

'HCOCOCOtJCW^IIOCOi— i«io« 

go 
faS5 

h-l                           1-4 

h- tl— 1                     HH 

PHOSPHORUS  METABOLISM 


453 


Children  Nos.  1,  2,  4  and  6  were  pathological  cases ;  Nos.  3  and 
5  were  healthy. 

Netter  (1900)  found  the  density  of  the  urine,  and  its  content  of 
nitrogen,  phosphorus  and  calcium  higher  with  artificially  fed  than 
with  normally  fed  infants.  The  amount  of  feces  was  greater  with 
the  infants  on  cow's  milk,  and  their  content  of  calcium  and  phospho- 
rus was  also  higher.  Netter  reported  balance  data  from  7 
infants  on  sterilized  cow's  milk  as  the  exclusive  diet.  A  part  of 
the  figures  follow. 

AVERAGE  DAILY  BALANCES  OF  NITROGEN,  CALCIUM  AND 

PHOSPHORUS  PER  KILOGRAM   BODY   WEIGHT   WITH 

HEALTHY  INFANTS  ON  STERILIZED  COW'S  MILK 

Grams 


N 

CaO 

P2O5 

No.  of 

Duration 

Milk 

Food 

Food 

Age 

Subject 

Weiarht 

in  days 

Urine 

Urine 

C.C. 

Balance 

Feces 
Balance 

Feces 
Balance 

Months 

0.297 

0.344 

0.004 

0.071 

0.198 

0.182 

1 

7270 

4 

168 

+0.199 

+0.095 

0.211 
0.006 
0.177 

+0.09 

0.342 
0.095 
0.197 

8 

2 

5860 

3 

137 

+0.303 

+0.027 
0.189 

+0.049 
0.307 

10.5 

0.004 

0.096 

0.16 

0.184 

3 

7310 

4 

122 

+0.133 

+0.024 

0.263 

0.007. 

0.229 

+0.025 

0.427 
0.057 
0.3 

10.5 

4 

4710 

4 

170 

+0.305 

+0.026 

0.236 
0.009 
0.125 

+0.068 

0.298 

0.06 

0.184 

7.5 

6 

8300 

3 

121 

+0.118 

+0.101 

0.248 
0.010 
0.16 

+0.052 

0.294 
0.096 
0.122 

9.0 

7 

6980 

3 

148 

+0.165 

+0.077 

+0.075 

7.5 

Netter  gives  as  the. average  gains  of  these  infants  the  follow- 
ing: 

Average  gain  Average  gain 

per  day  per  kilo  per  day 

Body  weight    . . . .  . 11.64 1.772 

Nitrogen    1.26 0.194 

CaO    0.395 0.058 

pa°«     0.402 0.059 


454  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Comparing  these  figures  with  results  of  Michel,  Netter  finds 
that  the  gain  in  nitrogen,  calcium  and  phosphorus  is  about  the  same 
as  for  new-born  naturally-fed  infants,  although  less  regular.  The 
gain  in  weight  is  said  to  be  less.  The  author  appends  118  referen- 
ces. 

P.  Miiller  (1900)  investigated  the  nucleins  and  lecithins  of  the 
feces  of  infants,  adults,  calves  and  dogs,  and  found  that  cow's 
milk  casein  in  the  intestine  of  the  normal  infant  leaves  no  more 
digestion-residue  rich  in  phosphorus  than  does  the  casein  of 
woman's  milk;  also  that  adults  utilize  the  casein  phosphorus  of 
cow's  milk  as  completely  as  do  infants.  Lecithin  was  found  in 
milk  feces  and  was  considered  as  derived  from  the  food. 

Von  Szontagh  (1902)  conducted  two  4-day  balance  experiments 
on  an  infant  with  "Szekely's  milk,"  a  modified  cow's  milk,  from 
which  the  casein  had  been  precipitated  by  liquid  carbon  dioxide. 
The  nitrogen  was  apparently  absorbed  to  the  extent  of  90.37-92.20 
percent  of  the  intake,  the  phosphorus  65.26-60.05  percent,  and  the 
calcium  10.79-15.33  percent.  The  retention  was  0.679-0.810  gm.  nitro- 
gen, 0.218-0.542  gm.  P,05  and  0.086-0.105  gm.  CaO. 

Cronheim  and  Miiller  (1903)  studied  the  effects  of  sterilization 
of  milk  on  its  utilization  by  infants.  The  results  on  phosphorus 
retention  were  inconclusive.  Later  studies  on  the  same  subject 
showed  that  sterilization  of  milk  is  probably  without  influence  on 
phosphorus  metabolism. 

Tangl  (1904)  conducted  two  four-day  balance  experiments  with 
infants  on  "Szekely's  milk."  The  composition  of  the  milk  was  said 
to  be  as  follows :  Water  87.2 ;  fat  3.7 ;  casein  1.5 ;  albumin  0.9 ;  milk 
sugar  6.3  and  ash  0.7  percent. 

The  milk  as  analyzed  for  the  two  experiments  was  composed 
as  follows : 

Analyses  of  Szekely's  Milk 

I  II 

Solids    13.13     . : 13.11 

Organic  matter 12.58    12.57 

Ash 0.555 0.534 

Total  N 0.293 0.344 

Protein   N    0.261 0.331 

Casein  N 0.235 0.307 

Albumin  N 0.026 0.024 

Non-protein  N 0.033 0.013 

Sugar    6.53     6.32 

Calories  in  100  gm 67.31     71.32 

K    0.163 0.163 

Na 0.031 0.032 

Ca 0.083 0.084 

Mg  0.0095 0.0097 

CI    0.103 0.084 

S  0.013 0.017 

P 0.069 0.074 


PHOSPHORUS  METABOLISM 


455 


The  apparent  absorption  of  N  was  90.0-92.3  percent  of  the 
intake.  The  retention  of  nitrogen  was  23.4-29.9  percent,  of  Ca 
11.5-14.2  percent,  and  of  P  18.6-38.5  percent  of  the  intake. 

Schlossmann  (1905a,  1905b)  says  that  during-  the  first  year 
of  an  infant's  life  it  retains  55-60  gm.  P.  He  shows  that  the 
phosphorus  content  of  artificial  milk  mixtures  is  not  likely  to  fall 
below  the  normal  for  human  milk.  He  submits  data  showing  that 
infants  absorb  much  more  phosphorus  from  mixtures  of  cow's  milk 
and  cream,  or  buttermilk  and  cream,  or  from  buttermilk  alone,  than 
from  woman's  milk. 

Bruck  (1908),  in  metabolism  experiments  on  artificially  nour- 
ished infants,  found  about  three  times  as  much  phosphorus  in  the 
urine  as  in  the  feces,  which  implies  satisfactory  absorption.  The 
numerical  data  are  as  follows. 

PATHS  OF  EXCRETION  OF  PHOSPHORUS  AND  THE  MINERAL  BASES 
WITH  INFANTS— Grams 


Exp.  No. 

Constituents 

Intake 

Urine 

Feces 

Percent  of 
retention 

CaO 

2.4098 

0.1944 

1.0229 

45.3 

I 
4  days 

MgO 

0.6392 

0.0269 

0.1815 

67.2 

P2O5 

2.8795 

1.7086 

0.5001 

23.5 

K2O 

2.7775 

1.2374 

0.4243 

40.2 

Na20 

1.7190 

0.7942 

0.0547 

50.6 

CaO' 

2.2276 

0.1094   ' 

1.2190 

40.3 

MgO 

0.3105 

0.0520 

0.0739 

59.5        l 

3  days 

P2O5 

4.3624 

2.5091 

0.9521 

26.3 

K2O 

3.9396 

4.2613 

0.1733 

Na20 

2.6684 

1.9060 

0.0143 

28.0 

The  diet  was  milk,  gruel  and  malt. 

Friedenthal  (1911)  reviewed  the  knowledge  on  the  ash  constitu- 
ents of  different  varieties  of  milk  in  connection  with  the  artificial 
feeding  of  infants.  A  table  gives  the  anions  and  cations  per  liter 
of  the  milk  from  woman,  cow,  sow,  dog,  rabbit  and  ass. 

These  comparisons  of  the  milk  of  the  various  species  of  animals, 
as  food  for  infants,  lose  somewhat  in  practical  significance,  as  also 
do  the  observations  on  proprietary  infant  foods,  and  the  old- 
fashioned  modification  of  milk  with  water,  since  the  modern  method 
of  "split-protein"  feeding,  which  is  based  on  the  modification  of 
milk  with  whey,  has  come  into  prominence.  Through  the  use  of 
skim  milk,  whey,  cream  and  milk  sugar,  all  from  cow's  milk,  an 
artificial  food  is  prepared  which  may  be  varied  in  the  details  of  its 
composition  in  such  ways  as  exactly  to  suit  the  requirements  of  any 


456  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

particular  infant.  This  system  has  made  possible  a  new  era  in 
successful  and  scientific  infant  feeding.  The  use  of  evaporated 
whey  in  powder  form  lessens  the  labor  incident  to  the  preparation  of 
the  milk  by  this  process.  The  advantages  of  this  system  are  (1) 
that  it  allows  of  the  control  of  the  proportions  of  fat  and  protein, 
(2)  that  it  makes  possible  the  increase  of  the  normal  proportion  of 
albumin  to  casein,  an  advantage  to  delicate  infants,  and  (3)  that  it 
assists  in  the  prevention  of  rachitis  and  other  disturbances  of  min- 
eral metabolism  in  infants,  by  providing,  through  the  use  of  whey, 
against  the  possibility  of  a  deficient  intake  of  calcium,  phosphorus 
and  other  mineral  nutrients  due  to  the  dilution  of  cow's  milk  with 
water  according  to  the  old  method,  and  also  against  excessive  outgo 
of  calcium  in  soap  stools. 

Some  additional  references  on  the  consideration  of  suitable  milk 
for  infant  feeding  are  listed  below :  Langgaard,  1875 ;  Biedert,  1874, 
1880,  1884,  1887,  1897 ;  Schmidt,  1882 ;  A.  V.  Meigs,  1883 ;  Schloss- 
mann, 1896;  Wroblewski,  1894a,  1894b,  1898;  Siegfried,  1897; 
Edlef sen,  1901 ;  W.  Camerer,  Jun.  and  Soldner,  1903. 

MISCELLANEOUS  DATA  ON  PHOSPHORUS  METABOLISM  OF  INFANTS 

Michel  (1897)  shows  that  of  all  the  constituents  of  the  mother's 
milk  the  minerals  are,  in  general,  least  well  utilized,  as  indicated  by 
comparison  of  food  and  feces.  The  phosphorus  was  utilized  in  a 
case  under  observation  to  the  extent  of  91.63  percent,  the  food  phos- 
phorus being  0.263  gm.  (P205)  in  566  gm.  milk,  and  the  feces  phos- 
phorus 0.022  gm.  in  2.99  gm.  dried  feces. 

Oechsner  (1899)  determined  the  nitrogen  and  phosphorus  in 
the  urine  of  nursing  infants.  The  24-hour  quantity  varied  from 
0.04  to  0.12  gm.  P205,  and  the  proportion  of  N:P205  from  5.2:1  to 
10.4:1. 

Keller  (1900a)  determined  that  the  organic  phosphorus  of  the 
urine  of  infants  was  greater  in  amount  after  feeding  cow's  milk 
than  after  feeding  human  milk,  though  it  was  a  smaller  percentage 
of  the  total  urinary  phosphorus,  since  the  total  phosphorus  from 
the  cow's  milk  greatly  exceeded  that  resulting  from  woman's  milk. 
It  varied  in  amount  from  less  than  one  to  about  10  percent  of  the 
total  urinary  phosphorus. 

Camerer,  Soldner  and  Herzog  (1902)  came  to  the  conclusion 
that  of  the  ash  constituents  given  in  the  first  month  of  the  life 
of  the  human  infant  (about  1.4  gm.  daily)  about  50  percent  is  used 
for  the  growth  of  the  body,  and  that  among  these  relatively  the 
largest  amounts  are  retained  of  lime,  magnesia  and  phosphoric  acid ; 
apparently  more  than  65  percent  of  the  amounts  given. 


PHOSPHORUS  METABOLISM 


457 


W.  Freund  (1905)  studied  the  influence  on  metabolism  of  vari- 
ation in  the  fat  content  of  the  milk  of  infants.  Increasing  the  fat 
increased  the  urinary  ammonia  coefficient,  decreased  the  feces  phos- 
phorus, and  increased  the  phosphorus  retention.  The  increased 
phosphorus  retention  was  explained  as  due  to  improved  absorption, 
a  part  of  the  calcium  ordinarily  excreted  combined  with  phosphorus 
in  the  feces  being  excreted  in  this  case  combined  with  fatty  acids, 
thus  leaving  a  corresponding  amount  of  phosphoric  acid  free  to  be 
absorbed. 

Michel  and  Perret  (1906)  report  figures  for  the  metabolism  of 
calcium  and  phosphorus  by  four  infants  four-and-a-half  to  five-and- 
a-half  months  of  age  showing  that  calcium  is  utilized  to  the  extent 
of  13.05-60.22  percent.  A  part  of  the  variation  in  the  apparent 
utilization,  as  above  stated,  is  due  to  variability  in  the  excretory 
function  of  the  intestine,  the  percent  of  utilization  being  deter- 
mined by  a  comparison  of  food  and  feces  only,  while  comparatively 
trivial  circumstances  may  deflect  katabolized  nutrients  from  urine  to 
feces. 

Heubner  (1909)  submits  the  following  data  showing  the  extent 
of  the  urinary  excretion  of  phosphorus  by  infants. 

PHOSPHORUS  EXCRETION  IN  THE  URINE  OF  HEALTHY  INFANTS 


Age 

Food 

Weight 
Kg-. 

Phosphorus  per  day 
per  kg-,  tody  weight 

Grams 

Phosphorus,  in 
percent  of  urine 

No.  of 
analyses 

3-5  weeks 

Mother's  milk 

3.16 

4.85 
3.86-4.38 
4.05-6.80 

5.23 
3.98-4.80 

0.0003 

0.0017 

0.0010 
0.0007 
0.0027 

0.0018 
0.0012 
0.0022 
0.0012 
0.0039 
0.0025 

2 

4 
5 

4 

Moll  (1909)  concluded  that  there  is  no  phosphorus,  or  at  the 
most  very  small  amounts,  in  the  urine  of  the  normal  breast-fed 
infant,  but  that  indigestion  causes  some  elimination  of  phosphorus 
by  this  channel.  Organic  phosphorus  in  the  urine  of  a  breast-fed 
infant  is  regarded  as  a  pathological  symptom. 

Heubner  (1910)  determined  the  phosphorus  (P)  of  the  urine  of 
the  new-born  infant  as  0.0018  percent,  and  the  phosphorus  of  the 
amniotic  fluid  as  0.003  percent.  These  values  are  about  the  same 
as  those  reported  by  other  investigators  from  blood  serum,  from 
which  Heubner  suggests  that  we  may  consider  0.002  percent  P  as 
the  physiological  concentration  of  body  fluids  in  soluble  phosphates, 
which  filter  out  into  the  urine,  and  that  this  is  the  minimum  value 
for  urine  content  from  this  source.  See  also  Langstein  and 
Memann  (1910). 


458  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Koeppe  (1911)  studied  urinary  phosphorus  elimination  in 
infants  as  affected  by  amount  and  kind  of  food,  and  by  NaCl  added 
to  the  food.  The  urinary  phosphorus  was  found  to  vary  with  the 
amount  of  food,  if  the  kind  remained  the  same,  and  with  the  kind, 
if  the  quantity  remained  the  same.  There  is  some  evidence  tending 
to  show  increased  concentration  of  phosphorus  in  the  urine  follow- 
ing the  ingestion  of  sodium  chloride,  but  the  results  are  hardly 
sufficient  to  establish  the  fact.  The  ingestion  of  NaCl  brought 
about  an  increase  of  temperature  and  a  chlorine  retention. 

When  infants  are  given  more  casein  in  the  milk  than  they  are 
able  to  digest,  especially  as  in  feeding  on  unmodified  cow's  milk, 
there  may  occur  in  the  feces  hard,  white,  or  yellowish  lumps  of 
undigested  curd,  having  much  the  same  appearance  as  the  lumps 
of  soap  in  the  feces  sometimes  resulting  from  an  excess  of  fat  in  the 
milk  (Monrad,  1911 ;  Uff  enheimer  and  Takeno,  1911 •;  J.  Bauer,  1911 ; 
Ibrahim,  1911;  and  Brennemann,  1911).  The  soap  and  curd  lumps 
are  commonly  distinguished  by  their  behavior  under  the  influence 
of  heat. 

Tobler  (1911)  published  studies  of  alterations  in  the  composi- 
tion of  the  muscles  of  infants  after  death  from  disorders  involving 
acute  or  chronic  loss  in  weight.  The  ash  content  of  the  muscles  of 
atrophic  children  was  very  low,  but  the  phosphorus  content  of  the 
ash  was  not  subnormal.  The  ash  of  the  fat-free,  dry  muscle  varied 
between  3.164  and  4.724  percent,  while  the  phosphorus  in  the  ash 
varied  between  7.16  and  14.63  percent.  Primary  water-loss  was 
considered  to  have  led  to  salt-loss.       See  also  Birk  (1911). 

E.  Muller  (1911)  thought  that  the  ill  effects  of  feeding 
infants  on  undiluted  cow's  milk  are  due  to  excess  of  whey  constitu- 
ents other  than  sugar,  and  submitted  balance  data  in  support  of  his 
idea.      This  is  not  the  prevailing  view. 

Hoobler  (1911)  shows  that  increasing  the  fat  of  the  food  of 
infants  increases  the  percentage  of  the  phosphorus  of  the  food 
which  is  absorbed.  With  milk  containing  2.1  percent  fat,  67 
percent  of  the  phosphorus  was  absorbed;  with  4  percent  milk  the 
phosphorus  absorption  was  78.1  percent,  while  with  5.4  percent  milk, 
83  percent  of  the  phosphorus  was  absorbed. 

Schloss  and  Crawford  (1911),  studying  phosphorus  metabolism 
in  the  new-born  infant,  found  a  marked  diminution  in  the  phospho- 
rus excretion  after  the  first  three  days,  there  being  an  inverse  rela- 
tion between  the  leucocyte  count  and  the  elimination  of  phosphorus 
and  uric  acid  during  this  period,  both  apparently  having  an  origin  in 
cell  nuclei. 


PHOSPHORUS  METABOLISM  459 

For  determinations  of  the  phosphorus  and  other  mineral 
elements  of  the  food  of  infants  and  children  see  Hoobler  (1912). 

G.  Wolff  (1912a)  conducted  metabolism  experiments  with 
infants  involving  quantitative  variation  in  the  diet.  He  concludes 
that  the  change  from  scanty  to  abundant  diet  produces  no  unfavor- 
able effect  on  the  metabolism  of  nitrogen,  phosphorus  and  calcium, 
and  that  calcium  and  phosphorus  are  likely  to  be  deficient  in  diets 
which  contain  nitrogen  sufficient  only  for  maintenance  or  slight 
storage. 

McCrudden  and  Fales  (1912),  studying  intestinal  infantilism, 
found,  in  two  cases,  poor  absorption  of  nitrogen,  sulphur,  phospho- 
rus, calcium  and  magnesium.  Calcium  was  excreted  in  the  feces 
in  amounts  greater  than  the  intake ;  the  other  elements  were  in  part 
retained. 

Jundell  (1913)  found  normal  calcium  and  phosphorus  absorption 
and  retention  (unlike  the  other  minerals)  with  2  children  suffering 
from  dyspepsia,  and  2  with  intestinal  intoxication.  This  result  is 
ascribed  to  the  effects,  on  the  solubility  of  the  tertiary  and  second- 
ary phosphates,  of  the  abnormal  amounts  of  fermentation  acids 
present. 

Giffhorn  (1913)  studied  the  effects  of  variations  in  the  fat 
and  in  the  whey  constituents  of  the  diet  of  infants  as  affecting  min- 
eral metabolism.  With  healthy  children  the  fat  in  the  diet  favored 
nitrogen,  phosphorus  and  magnesium  retention,  while  with  refer- 
ence to  calcium  the  results  were  inconclusive.  There  was  not 
noted  an  increased  mineral  retention  resulting  from  the  whey-rich 
diet  as  compared  with  one  containing  less  of  the  whey  constituents. 

Takeno  (1913)  studied  intestinal  elimination  by  children  in  18 
experiments  under  varied  dietary  conditions.  The  close  relation- 
ship of  calcium  and  phosphorus  was  apparent".  Phosphorus  elimin- 
ation was  lowest  on  nurse's  milk  diet,  and  comparatively  low  also 
on  milk  modified  with  whey ;  it  was  highest  on  a  mixed  diet.  Digest- 
ive disturbance  diminished  fecal  phosphorus  elimination.  The 
effects  of  cream  and  of  lipanin  on  phosphorus  elimination  in  the 
feces  was  doubtful  or  lacking,  while  cod-liver  oil  produced  a  marked 
effect  on  both  calcium  and  phosphorus. 

Kaminer  and  Mayerhofer  (1913)  concluded,  from  a  study  of  the 
urine  of  50  artificially  fed  infants,  that  the  metabolism  of  children 
so  nourished  is  not  normal,  that  there  is  regularly  a  slight  phos- 
phaturia,  and  that  this  phosphaturia  increases  with  intestinal 
indigestion  or  increasing  concentration  or  carbohydrate  content  of 
the  diet.  The  organism  may  then  adjust  itself  to  the  dietary 
change,  and  the  urinary  phosphorus  elimination  will  fall  again. 


460  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

For  phosphorus  requirements  of  infants  see  p.  414. 

Droge  (1913)  studied  the  effects  of  extirpation  of  the  blood 
glands,  the  testicles,  the  thyroid  and  especially  the  spleen  from 
suckling  dogs,  having  in  mind  Czerny's  idea  of  congenital  insuf- 
ficiency in  these  organs  as  a  cause  of  exudative  diathesis  in  human 
infants.  A  part  of  the  numerical  results  are  to  be  found  in  the 
discussion  of  the  thyroid  glands  in  relation  to  phosphorus  metabo- 
lism. A  certain  amount  of  modification  of  the  metabolism  of  the 
dogs  was  produced  by  the  removal  of  these  glands,  as  made  evident 
by  various  details  of  composition,  including  calcium  and  phosphorus 
content. 

Summary.  In  accord  with  the  relatively  slow  growth  of  the 
human  infant  the  ash  and  the  phosphorus  contents  of  woman's  milk 
are  low  in  comparison  with  the  milk  of  the  lower  animals.  It  has, 
however,  a  higher  content  of  phosphatid  and  nucleon  phosphorus,  a 
relatively  large  proportion  of  the  total  phosphorus  thus  being  in  an 
organic  form,  and  the  phosphorus  retention  from  woman's  milk  is 
greater,  in  proportion  to  the  intake,  than  from  cow's  milk. 

Because  of  the  higher  fat  and  casein  content  of  cow's  milk  than 
of  woman's  milk,  the  former,  when  fed  to  infants,  requires  much 
dilution,  which  reduces  the  calcium  and  phosphorus  more  nearly  to 
the  amounts  present  in  woman's  milk  and  reduces  the  lecithin  and 
nucleon  phosphorus  to  amounts  very  much  lower  than  those  found 
in  woman's  milk.  This  fact  has  been  considered  to  warrant  the 
early  introduction  of  egg-yolk  into  the  diet  of  the  artificially  fed 
infant. 

From  undiluted  cow's  milk  infants  may  absorb  and  retain  much 
more  phosphorus  than  from  woman's  milk,  but  the  ability  of  infants 
to  do  so  varies  much  with  the  individual. 

The  phosphorus  of  cow's  milk  appears  to  be  more  efficiently 
absorbed  than  the  calcium  of  the  same ;  more  total  and  organic  phos- 
phorus is  found  in  the  urine  of  infants  fed  on  cow's  milk  than  in 
the  urine  of  infants  receiving  the  mother's  milk ;  also  a  much  larger 
proportion  of  phosphorus  to  nitrogen  is  found  in  the  urine  from 
cow's  milk  feeding. 

Sterilization  of  milk  seems  to  be  without  any  such  effects  on 
phosphorus  metabolism  as  are  ascertainable  in  short  balance  experi- 
ments. 

The  modification  of  milk  with  whey,  instead  of  water,  is  in 
many  ways  advantageous,  and  provides  against  the  possibility  of 
deficiency  of  minerals.  The  feeding  of  excessive  quantities  of  whey 
solids,  made  possible  by  the  use  of  evaporated  whey,  has  been  known 
to  produce  oedematous  manifestations  in  delicate  infants,  apparent- 
ly through  the  superabundance  of  mineral  salts  provided. 


PHOSPHORUS  METABOLISM 


461 


There  is,  at  the  most,  but  very  little  phosphorus  in  the  urine  of 
normal  breast-fed  infants,  but  urinary  phosphorus  is  considerable 
on  a  mixed  diet,  and  is  increased  by  indigestion.  Organic  phospho- 
rus in  the  urine  of  breast-fed  infants  is  regarded  as  a  pathological 
symptom. 

There  is  a  diminution  in  the  urinary  phosphorus  and  uric  acid 
excretion  of  new-born  infants,  after  the  first  few  days,  and  a  coinci- 
dent inverse  variation  in  the  leucocyte  count. 

With  artificially  fed  infants  there  is  a  slight  phosphaturia 
which  increases  with  intestinal  indigestion  or  with  carbohydrates 
in  the  diet. 

Increasing  the  fat  of  the  food  increases  the  urinary  ammonia 
and  favors  phosphorus  absorption  and  retention. 

Absorption  of  the  minerals  is  deficient  in  infantilism,  but  in 
intestinal  indigestion  and  in  dyspepsia  the  calcium  and  phosphorus 
absorption  and  retention  may  be  normal. 

For  further  information  on  this  matter  see  Phosphorus  Require- 
ments of  Infants,  pages  414-419. 

EFFECTS  OF  MENTAL  WORK  ON  PHOSPHORUS  EXCRETION 

In  view  of  the  vividness  of  ones  consciousness  of  his  own  mental 
operations  it  becomes  a  matter  of  interest  to  consider  the  extent  of 
the  metabolic  accompaniments.  One  does  not  readily  accept, 
without  question,  the  dictum  that  intense  mental  effort  produces  no 
recognizable  end  products. 

Byasson(1868)  studied  urine  composition  as  affected  by  mental 
work,  rest  and  exercise,  the  diet  being  controlled  by  analysis.  The 
P205  of  the  urine  during  rest  was  1.508  gm.  per  day,  during  muscu- 
lar activity  1.4779  gm.  and  during  cerebral  activity  1.9777  gm.  These 
differences  in  outgo  are  not  characteristic  of  the  results  of  later 
work.      See  also  Wood,  L.  H.,  (1869). 

Speck  (1882)  failed  to  determine  an  increase  of  oxidation,  as  a 
result  of  mental  work,  by  examination  of  the  inspired  and  expired 
air. 

Mairet  (1884b)  studied  metabolism  as  affected  by  mental  work. 
A  part  of  his  data  are  the  following: 

TWENTY-FOUR  HOUR  URINE  DATA  AS  AFFECTED  BY  MENTAL  WORK 

Grams 


Mixed  diet 

Vegetable  diet 

Low  diet 

Condition 

Nitrosren 

Alkali 

phosphoric 

acid 

Nitrosren 

Alkali 

phosphoric 

acid 

Nitrosren 

Alkali 

phosphoric 
acid 

24.54 
22.00 

1.65 
1.53 

10.82 
8.45 

1.16 
1.10 

12.13 
10.71 

1.13 

0.99 

462 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


We  see  here  no  evidence  of  increased  metabolism  as  a  result  of 
mental  work. 

Schtscherbak  (1890)  studied  phosphorus  metabolism  as  affect- 
ed by  mental  work.  The  results  do  not  warrant  conclusions.  The 
author  states  that  he  found  the  venous  blood  from  the  brain  to 
contain  0.07-0.12  parts  per  1000  less  P205  than  the  arterial. 

Preysz  (1891,  1892-3)  found  no  increase  of  urinary  phosphorus 
after  mental  exercise. 

Mainzer  (1902)  reviewed  the  literature  of  the  subject  of 
metabolism  as  affected  by  mental  activity,  and  at  a  later  date  (1903) 
reported  on  studies  of  metabolism  during  fast,  and  as  affected  by 
mental  work,  and  protracted  wakefulness.  Urinary  analyses  only 
were  made.  Mainzer  considered  the  curves  of  elimination  of 
nitrogen  and  phosphorus,  as  affected  by  mental  work  and  by 
protracted  wakefulness,  to  be  similar,  both  being  fatigue  curves. 

Gilbert  and  Posternak  (1903)  determined  in  metabolism  experi- 
ments that  neither  prolonged  brain  activity  nor  extreme  nervous 
excitement  produces  any  certain  effect  on  urinary  phosphorus  elim- 
ination. Such  variations  as  were  noted  seem  to  be  within  the 
normal  range. 

Aron  and  Hocson  (1911a)  studied  nitrogen  and  phosphorus 
excretion  in  the  urine  as  affected  by  mental  work.  The  results  as 
submitted  are  inconclusive. 

DAILY   NITROGEN  AND   PHOSPHORUS   EXCRETION   IN   THE   URINE 
DURING  PERIODS  OF  REST  AND  MENTAL  WORK— Grams 


Condition 


Subject 


P2O5 


Rest,  2  days 

Mental  work,  2  days 

Rest,  2  days 

Rest,  2  days 

Mental  work,  2  days 

Rest,  3  days,  1st  day 

Rest,  3  days,  2nd  day 

Rest,  3  days,  3rd  day 

Mental  work,  3  days,  1st  day.. 
Mental  work,  3  days,  2nd  day. 
Mental  work,  3  days,  3rd  day. 

Rest,  2  days,  1st  day 

Rest,  2  days,  2nd  day 


Prisoner  D 

H 

E' 


11.01 
12.96 
11.41 
13.75 
13.77 
9.09 
8.80 
9.01 
8.66 
8.41 
9.13 
8.45 
8.76 


1.42 
1.85 
1.56 
2.14 
2.42 
1.65 
1.13 
0.91 
1.08 
0.98 
1.28 
1.29 
1.25 


Voit  (Hermann's  Handbuch)  determined  long  ago  that  the  total 
phosphorus  content  of  the  nervous  system  of  an  adult  human  being 
is  about  12  gm.  P205,  of  the  muscles  about  130  gm.  and  of  the  bones 
about  1400  gm.  With  these  figures  in  mind,  and  considering  the 
normal  variability  of  the  phosphorus  outgo  under  apparently 
constant  conditions,  and  the  fact  that  the  brain  is  not  a  highly 
vascular  organ,  one  would  not  be  led  to  anticipate  marked  or  even 


PHOSPHORUS  METABOLISM 


463 


appreciable  effects  of  brain  work  on  total  phosphorus  outgo.  At 
the  same  time,  however,  we  must  note  that  these  experimental  data 
do  not  prove'  that  there  is  no  increased  katabolism  of  the  brain 
tissue  during-  mental  work.  They  simply  do  not  prove  that  there  is 
such  increase. 

PHOSPHORUS  METABOLISM  AS  AFFECTED  BY  NERVE  STIMULATION 

Weyl  and  Zeitler  (1882)  studied  the  composition  of  rabbit's 
muscle  as  affected  by  electrical  stimulation,  with  especial  reference 
to  the  acid  reaction  developed.  They  state  the  supposition  that  the 
process  is  a  union  of  phosphoric  acid,  resulting  from  katabolic 
decomposition,  with  K2HP04  to  form  2KH2P04.  In  an  effort  to 
learn  the  source  of  the  free  phosphoric  acid  involved  in  the  reaction 
Weyl  and  Zeitler  estimated  the  total  and  lecithin  phosphorus  in  rest- 
ing muscles  and  in  others  stimulated  for  30-50  minutes.  Fresh 
muscle  they  found  to  contain  (7  determinations)  0.65-0.82  percent 
of  lecithin,  and  stimulated  muscle  (3  determinations)  0.62-0.665 
percent.  Inorganic  phosphates,  however,  increased  consistently 
and  to  a  marked  extent.      A  part  of  the  data  are  as  follows : 

PHOSPHATE  AND  LECITHIN  CONTENT  OF  RABBIT  MUSCLE  AS 
AFFECTED  BY  STIMULATION 


Experiment 

State  of  muscle 

Length,  of 
stimulation 

Minutes 

Weight  of 

muscle  in 

grams 

Inorganic 
phosphates 

Percent 

Lecithin 
Percent 

I 

Resting 
Stimulated 

30 

72 
75 

0.308 
0.357 

0.669 
0.665 

II 

Resting" 
Stimulated 

45 

54 

53 

0.262 
0.318 

0.707 
0.623 

III 

Resting1 
Stimulated 

50 

71 

65 

0.341 
0.362 

0.816 

IV 

Resting: 
Stimulated 

45 

79 
71 

0.288 
0.334 

0.709 
0.655 

Since  the  lecithin  phosphorus  decrease  does  not  account  for  the 
inorganic  phosphate  increase  the  authors  conclude  that  this  increase 
of  phosphates  takes  place  at  the  expense  of  the  nucleins — which 
were  not  estimated. 

Krchivetz  (1900)  investigated  phosphorus  elimination  as 
affected  by  nerve  stimulation  in  rabbits,  but  the  results  seem  to  us 
to  be  of  doubtful  significance.  The  same  may  be  said  of  the  experi- 
ments of  Malerba  (1905),  who  studied  the  phosphorus  compounds  in 
both  venous  and  arterial  blood  from  the  brain  of  the  dog,  before  and 
after  subjection  to  pain.     No  important  differences  were  observed. 


464  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

The  function  of  inorganic  phosphate  in  the  physiology  of 
striated  muscle  is  explained  by  E.  B.  Meigs  (1912)  in  brief  as 
follows:  Stimulation  of  muscle  causes  the  production  of  lactic 
acid,  the  presence  of  which  in  the  muscle  fibers  causes  the  sarco- 
styles  to  swell  at  the  expense  of  surrounding  sarcoplasmic  spaces, 
the  shortening  of  the  muscle  resulting  from  the  increased  volume  of 
its  sarcostyles.  The  relaxation  of  the  muscle  is  brought  about  by 
the  combination  of  the  lactic  acid  with  the  potassium  phosphate 
contained  in  the  muscle  fibers. 

See  also  the  work  of  Siegfried  and  of  Cavazzani  on  phospho- 
carnic  acid  metabolism  p.  303-305. 

EFFECTS  OF  EXERCISE  ON  PHOSPHORUS  METABOLISM 

Pettenkofer  and  Voit  (1866)  reported  results  of  complete  food 
studies  on  man  by  metabolism  and  respiration  experiments.  In 
connection  with  this  work  urinary  phosphorus  estimations  were 
made  which  showed  that  with  a  medium  diet  urinary  phosphorus 
was  not  increased  by  work. 

Flint  (1871)  studied  metabolism  as  affected  by  protracted  walk- 
ing. The  urinary  excretion  of  phosphorus  was  much  greater  during 
the  period  of  walking  than  in  fore-  and  after-periods  of  moderate 
exercise.  During  the  walking  days  the  phosphorus  outgo  varied 
almost  as  the  distance  covered. 


P2O5  of  urine 

Walking1  day 

Miles  walked 

Grams 

3 

92 

6.625 

1 

80 

5.504 

4 

57 

4.296 

2 

48 

4.674 

5 

40.5 

3.725 

Engelmann  (1871)  in  studying  the  effects  of  exercise  on  metabo- 
lism concluded  that  no  parallelism  exists  in  the  excretion  of  urea, 
phosphoric  acid  and  sulphuric  acid.  The  food  and  water  ingested 
were  controlled,  and  amounts  kept  constant.  The  exercise  consisted 
of  walking,  running,  hill  climbing,  digging,  chopping  and  sawing, 
and  was  carried  to  the  point  of  fatigue.  Great  thirst  was  experi- 
enced during  periods  of  activity.  The  experiments,  three  in  number, 
were  6-8  days  in  length,  half  of  each  experiment  being  spent  at 
exercise  and  half  at  rest.  There  were,  in  two  cases  out  of  three, 
slight  increases  in  the  urinary  excretion  of  phosphorus  and  sulphur 
during  work.  In  the  third  case  the  urine  and  feces  phosphorus 
together,  in  the  work  period,  slightly  exceeded  the  same  during  rest, 
though  urine  phosphorus  alone  was  slightly  lower  during  work. 


PHOSPHORUS  METABOLISM  465 

Penzoldt  and  Fleischer  (1882)  studied  urinary  phosphorus 
excretion  in  dogs  as  affected  by  dyspnoea,  etc.  Dyspnoea,  with  the 
attendant  increased  muscular  work,  caused  increased  phosphorus 
outgo,  followed  afterward  by  such  decrease  as  left  average  figures 
without  great  change.  The  lack  of  oxygen  alone  caused  an  increased 
phosphorus  outgo  which  persisted  into  the  after-period ;  while 
apnoea  caused  a  decreased  phosphorus  outgo  followed  afterward  by 
a  strong  increase.     See  also  Speck  (1882) . 

Pavy  (1876a,  1876b,  1877)  made  studies  of  the  urine  of  E.  P. 
Weston,  as  affected  by  protracted  walking.  While  there  was  appar- 
ently a  great  increase  in  phosphorus  elimination  as  a  result  of  the 
exercise,  there  were  no  phosphorus  determinations  on  either  food  or 
feces. 

North  (1883-4)  concluded,  as  a  result  of  experiments  on  him- 
self, that  phosphorus  elimination  is  not  increased  by  work,  unless 
the  labor  be  severe.  In  three  experiments  the  results  showed,  on 
the  whole,  no  important  effects  on  nitrogen  or  phosphorus  outgo. 

Mairet  (1884a)  studied  urinary  phosphorus  elimination  as 
affected  by  exercise  and  mental  work.  With  exercise,  on  a  meat 
diet,  the  urinary  phosphorus  was  not  changed ;  on  a  mixed  diet,  it 
was  increased  from  2.11  to  2.27  gm.,  and  on  a  vegetable  diet  from 
2.03  to  2.37  gm.  In  the  second  case  the  alkali  phosphates  increased 
from  1.57  to  1.74  gm.,  and  in  the  third  from  1.52  to  1.92  gm.  Urinary 
nitrogen  was  also  increased  by  exercise. 

Preysz  (1891,  1892-3)  determined  his  own  daily  urinary  phos- 
phorus excretion  during  10  days.  The  maximum  excretion  was  3.00 
gm.,  minimum  2.56,  mean  2.784  gm.  P205.  During  the  eleventh  day 
exercise  was  taken,  and  the  phosphorus  excretion  rose  to  4.17  gm. 
A  return  to  the  method  of  life  of  the  first  10  days  reduced  the 
urinary  phosphorus  outgo  to  2.616  (mean).  Further  tests  gave 
concordant  results. 

Klug  and  Olsavszky  (1893)  studied  urinary  phosphorus  elimin- 
ation with  a  dog  as  affected  by  exercise  and  by  lactic  acid  intake. 
The  average  outgo  for  a  10-days  preliminary  period  of  inactivity  was 
0.3175  gm.,  the  largest  amount  being  0.39  gm.,  and  the  smallest  0.24 
gm.  During  one  day  of  activity  the  excretion  was  0.57  gm.,  and  in 
the  next  day  of  inactivity  0.28  gm.  This  increase  of  phosphorus 
excretion  in  activity  the  authors  believe  to  be  due  to  the  solvent 
effect  of  the  lactic  acid  formed.  In  a  direct  test  of  the  effect  of 
lactic  acid  ingestion  on  phosphorus  elimination  the  results  are  incon- 
clusive, though  they  would  be  decisive  in  showing  increased  urinary 
phosphorus  with  lactic  acid  ingestion  were  it  not  for  the  data 
representing  the  fore-period. 


466  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

I.  Munk  (1895)  studied  the  effect  of  exercise,  on  a  constant  diet, 
on  the  urinary  excretion  of  nitrogen,  sulphur,  phosphorus,  calcium 
and  potassium.  Calcium  was  also  determined  in  the  feces.  An 
increased  excretion  of  phosphorus  as  a  result  of  exercise  was  clearly 
shown.  The  evidence  as  to  increased  calcium  excretion  is  less 
satisfactory.  Munk  considers  that  the  data  show  a  certain  amount 
of  decomposition  of  bone  tissue  as  a  result  of  work. 

Dunlop,  Paton  and  Stockman  (1897-8)  investigated  the  subject 
of  the  influence  of  exercise,  sweating  and  massage  on  metabolism 
generally.  In  connection  with  this  study  observations  were  made 
on  phosphorus  excretion.  These  authors  show  that  excessive 
muscular  work  causes  an  increased  phosphorus  outgo,  apparently 
from  the  katabolism  of  muscle  substance,  and  that  with  individuals 
out  of  condition  there  may  be  a  more  deep-seated  waste  of  tissue, 
involving  the  nucleins,  as  indicated  by  increased  excretion  of  uric 
acid.  Sweating  and  massage  were  found  without  influence  on 
phosphorus  excretion.  Thus  exercise  causes  katabolic  changes  in 
the  nitrogenous  tissues  of  persons  out  of  condition  that  are  not 
produced  in  the  bodies  of  men  in  training. 

Garratt  (1898)  investigated  metabolism  as  affected  by  exercise 
and  Turkish  baths.  The  exercise  consisted  of  bicycle  rides,  3  of  80 
miles,  one  of  71,  one  of  47  and  one  of  41  miles.  The  subject  was  in 
good  physical  condition,  and  the  fatigue  induced  was  never  excess- 
ive.     The  diet  was  of  normal  mixed  character,  but  was  not  analyzed. 

This  exercise  caused  an  excretion  of  urea  rising  to  a  maximum 
of  possibly  double  the  normal  in  about  12  hours,  and  only  regaining 
the  usual  level  after  30  hours  or  more  had  elapsed,  this  increase 
beginning  immediately  after,  but  not  during  the  exercise. 

Uric  acid  excretion  was  also  increased,  possibly  during  and 
certainly  immediately  after  the  exercise,  the  height  of  the  rise,  and 
duration  of  the  increase  being  less  with  a  subject  in  good  condition, 
and  with  sufficient  food,  than  with  a  subject  out  of  condition,  or  on 
insufficient  food.  Phosphorus  elimination  was  increased,  not  during 
the  exercise  but  immediately  after. 

Sulphate  excretion  was  also  increased,  the  rise  commencing 
during  the  exercise,  and  reaching  a  maximum  of  perhaps  3  times  the 
normal  within  six  hours. 

The  Turkish  baths  were  without  notable  effect  on  phosphate, 
uric  acid,  urea  or  sulphate  excretion,  but  caused  a  reduction  in  the 
excretion  of  chlorides. 


PHOSPHORUS  METABOLISM  467 

The  data  submitted  include  tables  showing-  the  effects  of  ordin- 
ary meals,  and  of  an  isolated  meal,  on  urinary  elimination.  The 
variations  due  to  meals  seem  quite  considerable  in  comparison  with 
those  noted  as  results  of  exercise.  Accurate  control  of  the  food 
consumed  during  the  exercise  experiments  would  have  been 
desirable. 

In  connection  with  the  above  observations  as  to  the  effects  of 
Turkish  baths  we  cite  a  pathological  case  of  excessive  perspiration. 
Marischler  (1898)  describes  the  case  of  a  boy  of  14  who  since  the 
age  of  8  years  had  perspired  profusely  at  all  times,  especially  in  cold 
weather.  By  balance  experiments  there  was  found  to  be  a  loss  of 
nitrogen,  calcium  and  phosphorus  from  the  body.  The  nitrogen 
and  phosphorus  of  the  urine  were  as  100:20  and  of  the  urine  and 
feces  together  100:25.  Atropin  lessened  the  perspiration,  but 
increased  the  phosphorus  elimination. 

Muscle  analyses  reported  by  Macleod  (1899)  furnish  some 
evidence  as  to  the  chemistry  of  muscular  activity,  in  particular  with 
regard  to  Siegfried's  theory  as  to  the  value  of  the  nucleon  of  muscle. 
Macleod  compared  the  distribution  of  the  phosphorus  in  the  muscles 
of  the  legs  of  dogs  that  had  been  exercised  in  a  treadmill  for  several 
hours  before  being  killed  with  that  in  the  corresponding  muscles  of 
dogs  that  had  been  resting  at  least  twelve  hours  before  being  killed. 
Determinations  were  made  of  the  moisture,  total  phosphorus,  phos- 
phorus soluble  in  water,  inorganic  phosphorus  in  the  water  extract, 
and  the  phosphorus  and  nitrogen  of  the  iron-nucleon  precipitate. 
From  these  values  were  also  computed  the  organic  phosphorus  of  the 
water  extract,  and  the  portion  of  this  organic  phosphorus  which  was 
not  in  the  form  of  nucleon.  The  results  did  not  support  the  theory 
so  far  as  the  nucleon  is  concerned.  There  was  a  large  loss  of  the 
organic  phosphorus  of  the  water  extract,  but  less  from  the  nucleon 
than  from  the  other  portion.  The  inorganic  phosphates  of  the 
water  extract  showed  corresponding  increase,  making  it  appear  that 
some  organic  phosphorus  compound,  by  its  decomposition,  plays  an 
important  part  in  muscle  metabolism  during  exercise.  It  is  to  be 
noted  also  that  the  total  phosphorus  of  the  muscle  apparently 
increased  during  the  exercise  by  an  increase  in  the  compounds  not 
soluble  in  water.      (See  tables  on  p.  468). 

Macleod  now  interprets  these  results  (1914,  personal  interview) 
as  due  first  to  a  formation  of  lactic  acid,  and  second  to  a  decomposi- 
tion, by  this  acid,  of  organic  phosphorus  compounds. 


468 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


o 

o  , 

to  .. 

20*0, 

01 

Ctf 

s 
rt 
> 
•u 

■-  ^  a  ©  s    ■*< 
Jllll   W 

i-H        COCOOOO 
HMOOHO 

2 

(4 

ft-S^ 

at*?'-    ^ 

OSlOSOCMiJtOS 

co^xcocmcmcni 

CO 

iHl-il-H  — -li-Hi-l 

TH 

*>0 

a * 

«  a 

XI  o 

5-^u 

•E^'pm 

OS  00  T  t—  CM  OS 

CO 
OS 

cm  coco -win  co 

CO 

a 

£8^ 

V> 

£ 

iocotjicocmco 

CO 

o 

r   O  +J 
IS 

rHlHlHlHt-HlH 

3 

.y  £•§  ap 

CONlOO'*'* 

■»JI 

w 

OOOOOO 

CO 

o 

SSooQ 

o'oo'ocso 

o 

°.S£ 

cor^^r^cMT* 

CO 

<S 

!Z5 

iMOOOOO 

o 

oooooo 

o 

a 

M 

u 

a 

14 

CO  CO  OS  CO  CO  OS 
CM  CO  CM  COT*  CM 

CO 

fe 

fc 

o 

oooooo 

o 

a 

rt 

a 

n.8    ^ 

?a    w 

i-nri-HCOO"£>. 

r- 

u 

H 

i-HOi-HOOO 

o 

— 

oooooo 

o 

3 

a 

>, 

P4 

u 

•Hn 

osoo^oor^co 

•^■n.  CD  OS  CO  CO 

$ 

a  i 

OOOOuO 

a 

Q 

Pra 

OOOOOO 

o 

ta 

H 

o 

o 

u 

CM 

u 

+S 

1 

ooosiracoojg 

co 

O 

In 

i-H  CM  CM  CM  CM  i-t 

CM 

s» 

oooooo 

o 

o 
a 

* 

CM 

cocposoos-n 

CM  CM  CM  CO  CM  CM 

ITS 

H 

S 

CM 

oooooo 

o 

H 

«4 

OSCMOOSt^cM 

intooiT-icoco 

CO  CO  ■**•"»<  CO  CO 

CO 

co 

OOOOOO 

o 

H 

s 

U   U    fc  L|   U   U, 

< 

cm  t-»-^  corpus 

l~l 

j 

1 

12 

iHCMCO^lOCO 

a 

rt 

J 

«-l 

o 

a  A  a^'^ 

3 

3 
rt 

Organic 
water  e 

tract  mi 
nucleon 
nucleon 

H/F 

ITS 

CMOS        CD 

•^co^;o 

HHrtri 

1-1 

CM 

■p 

a 

(4 

t 

o 

.S.S 

.H  rt  <J  rt      63 

COOS     'CM 
•     .CO     . 

com-Hio 

t~ 

u  o  s  M      M 

i-l  i— li-Hr-< 

rt 

a rt 

O-"^ 

■    •■ 

tu  3 

.Sf^n, 

ja  o 

6*-    <j 

CO 

£,n  vi     < 

CM  CM  CM  CO 

o 

«2%     H 

>   O-P 
P-  HI 

1— ItHiH-H 

1-t 

a 

o 

w 

o 

s 

0-<5HOSIM 
COCOCO-<Sl 

co 

is 

■q 
Eh 

MS   w 

1H 

1H 

tf 

m 

w 

H 

fa 

w 

■g|  a  §5 

COOIOCO 
O-^OCM 

OOOO 

■H 

0 

03 

8MB 

oooo 

0 

o 

°.Sifl 

o 

Q 

CO 

fa 

cS 

^5 

oo    -o 

ITS 

0 

o 

a 

oo    -o 

0 

03 

fa 

1-1 

^ 

03 

COCO^»-t 

fe 

Ah 

0 

oooo 

0 

u» 

a 

s 

rt 

a 

5 
u 

CM^-lt— O 

CO  OS  OS  CM 

9 

03 

u 

CO 

w 

P  a    ^ 

1— ll— 1<— 11— 1 

p 

a 

OS 

o 

w 
fa 

03 

o 

w 

fa 

a 

o 
o 

fi 

o 

r^cjoeo 

CO  ^1  CM  TT 

oooo 
oooo' 

8 

0 
0 

o 

PU 

fa 
o 

+1 

a 

5SSSS5 

31 

o 

rt 

CMIMCNJCM 

03 

M 

u 

oooo 

0 

fa* 
o 

4-> 

rt 

a 

M 

Pm 

fa 

r-  os  co  «5 

g 

n 

4J 

O 

CS  CM  CM  CM 
OOOO 

CM 

0 

fc 

CM  CO  .HO 

co  co  co  00 

^f( 

<! 

>tf1  ^tl  V  CO 

1»l 

o 

oooo 

0 

Eh 

■      ^   . 

s 

ftilK 

< 

COCO 

B  u 

1 

* 

^i 

2   1 

PHOSPHORUS  METABOLISM 


469 


I.  Kaup  (1902)  concluded  that  there  was  no  increased  protein 
metabolism  because  of  exertion.  There  were  no  food  phosphorus 
figures.  The  author  states  that  moderate  exercise  even  resulted  in 
phosphorus  retention. 

Maillard  (1908a,  1908b,  1909)  studied  urinary  phosphorus 
elimination  by  ten  soldiers  during  6  days,  on  an  ordinary  mixed  diet, 
the  object  being  to  determine  the  effects  of  muscular  exercise  and 
the  use  of  wine.  No  effects  of  the  use  of  wine  could  be  noted,  on 
the  nitrogen  and  phosphorus  elimination;  but  the  exercise,  which 
was  arranged  to  increase  in  severity  from  day  to  day,  in  two  three- 
day  experiments,  caused  an  undoubted  increase  in  phosphorus 
elimination  in  the  urine.  The  total  acidity  of  the  urine  increased, 
as  well  as  the  undetermined  nitrogen,  and  also  to  a  slight  extent 
the  uric  acid;  while  the  urea  decreased  a  little,  and  the  ammonia 
nitrogen,  purin  bases,  nitrogen  precipitable  by  silico-tungstic  acid, 
and  total  nitrogen  were  unchanged.  The  following  figures  show 
the  effects  of  exercise  on  phosphorus  elimination  in  the  urine. 


Exercise 

1st  d  ay- 
Light 

2nd  day- 
Medium 

3rd  day 
Severe 

4th  day 
Light 

5th  day 
Medium 

6th  day 
Severe 

P2O5 
P:N 

1.76 
1:48.4 

1.72 
43.3 

2.54 
33.1 

1.91 
44.4 

2.25 
35.4 

2.52 
33.4 

Scaffidi  (1910-11)  studied  purin  metabolism  as  affected  by  the 
fatigue  of  mountain  climbing.  A  portion  of  his  phosphorus  figures 
follow : 

DAILY   PHOSPHORUS   BALANCES   AS   AFFECTED   BY   FATIGUE   OF 
CLIMBING  HIGH  MOUNTAINS— Grams 


Intake 

Urine 

Feces 

Urine  and  feces 

Balance 

Balance 

Day 

P2O5 

P2O5 

P2O5 

P2O5 

P2O5 

N 

1 

3.634 

2.29 

0.58 

2.87 

+0.764 

+2.45 

2 

' ' 

2.42 

0.37 

2.79 

+0.844 

+3.149 

3  Fatigue 

2.712 

0.96 

3.67 

-0.036 

-2.001 

4 

2.938 

0.48 

3.41 

+0.224 
+0.544 

+2.039 

5 

2.75 

0.34 

3.09 

+1.089 

6 

2.54 

1 

2.70 

1.900 

0.48 

2.38 

+0.32 

+2.055 

2 

*  * 

1.593 

0.34 

1.93 

+0.77 

+4.525 
+2.190 

3  Fatigue 

2.363 

0.919 

0.672 

1.839 

+0.40 

4 

2.70 

1.995 

1.37 

2.93 

-0.23 

+3.155 

5 

1.870 

0.725 

2.31 

+0.39 

+3.915 

1 

3.237 

1.89 

0.78 

2.67 

+0.567 

H 

1-2.019 

2 

' ' 

1.84 

0.24 

2.08 

+1.157 

- 

-5.069 

3 

2.62 

0.25 

2.87 

+0.367 
+0.159 

- 

-3.239 

4  Fatigue 

2.328 

0.75 

3.078 

-4.439 

5 

2.88. 

1.25 

4.23 

-0.993 

-3.489 

1          6 

2.14 

0.64 

2.78 

+0.457 

!-3.219 

470  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Scaffidi  concluded  that  during  muscular  activity  there  is  no 
increase  of  uric  acid  elimination,  though  there  is  a  slight  increase 
of  purin  base  elimination ;  also  as  follows : 

Immediately  after  ending  the  muscular  work,  there  is  a  more 
or  less  pronounced,  but  always  quickly  developing  and  very  evident 
increase  in  the  amount  of  uric  acid  eliminated.  During  the  same 
time  the  purin  base  elimination  falls.  The  increase  of  uric  acid 
elimination  is  more  pronounced  if  the  work,  and  especially  the  rest 
following,  is  not  taken  at  too  high  an  altitude. 

The  total  purin  nitrogen  always  rises  on  the  working  day ;  often 
this  increase  continues  also  for  two  days  following  the  work.  This 
increase  is  due  almost  entirely  to  the  increased  formation  of  uric 
acid,  which,  as  may  be  concluded  from  the  phosphorus  content, 
comes  also  from  purin  bases  which  result  from  a  tearing  down  of 
nucleoproteids. 

The  elimination  of  phosphorus  rises  considerably  above  the 
normal  mean  on  the  working  days  and  on  the  following  days. 

The  total  elimination  of  nitrogen  is  not  much  changed  from  that 
of  the  resting  days,  if  one  leaves  out  of  account  the  cases  when  the 
work  undertaken  was  too  great,  because  of  lack  of  previous  training, 
or  because  it  was  carried  to  excessive  duration  or  intensity. 

The  ammonia  content  of  the  urine,  and  still  more  the  acidity, 
rise  in  consequence  of  labor. 

See  also  Hammond  (1863)  and  Lepine  and  Jacquin  (1879). 

Summary.  Ordinary  activity  does  not  cause  appreciable  increase 
in  phosphorus  outgo,  nor  does  more  vigorous  exercise  of  a  man  in 
training.  Severe  exercise,  however,  carried  to  the  point  of  tissue 
destruction,  and  this  is  by  no  means  a  rare  condition  in  the  lives  of 
many  people,  may  cause  marked  increase  in  the  outgo  of  phosphorus, 
uric  acid  and  other  tissue  katabolites.  The  principal  part  of  the 
increased  phosphorus  outgo  occurs  after  the  termination  of  the 
exercise,  and  the  supernormal  outgo  may  persist  during  several 
days.  The  phosphorus  outgo  is  not  increased  by  sweating  or 
massage. 

The  increased  phosphorus  outgo  after  severe  exertion  is  with- 
out doubt  due  to  tissue  destruction,  the  muscles  probably  contrib- 
uting at  least  the  principal  part. 

PHOSPHORUS  METABOLISM  DURING  PREGNANCY 

Hugounenq  (1899a)  studied  the  retention  of  minerals  by  the 
human  fetus.  The  retention  of  minerals  by  the  fetus  is  slight  at 
first,  but  very  active  at  the  end.  At  birth  the  infant  contains  about 
100  gm.  of  salts.  During  the  last  three  months  of  gestation  the 
fetus  acquires  twice  as  much  mineral  matter  as  previously. 


PHOSPHORUS  METABOLISM  471 

Ver  Eecke  (1900)  studied  metabolism  as  affected  by  pregnancy 
in  rabbits.  There  was  in  each  of  16  cases  a  very  marked  decrease 
in  phosphorus  elimination  during-  the  last  week  of  pregnancy,  to  a 
figure  usually  below  that  for  the  period  of  sexual  inactivity,  or  for 
the  first  week  of  pregnancy.  Between  the  first  and  last  weeks  of 
pregnancy,  that  is,  during  the  gestation  period,  the  phosphorus 
excretion  was,  in  twelve  cases  out  of  16,  higher  than  during  either 
the  first  or  the  last  week.  The  author  connects  the  decrease  in 
phosphorus  outgo  with  increase  in  activity  of  the  mammary  glands. 

Jagerroos  (1902),  studying  nitrogen  metabolism  in  pregnancy, 
with  dogs,  reaches  the  conclusion  that  in  this  condition  there  is  a 
marked  tendency  toward  parallelism  in  nitrogen  and  phosphorus 
metabolism,  as  is  natural,  considering  the  fact  that  the  pregnant 
female  is  in  a  sense  a  growing  animal,  and  also  that  there  is  usually, 
in  the  first  half  of  the  pregnancy,  a  period  of  increased  protein 
decomposition. 

Michel  (1899)  shows  the  increasing  demand  of  the  developing 
human  fetus  for  phosphorus.  The  figures  are  to  be  found  on  p. 
110. 

Schkarin  (1910)  fed  female  dogs  on  various  diets,  vegetable  and 
animal,  and  determined  the  total  phosphorus  of  the  pups  at  birth, 
and  after  suckling  for  varying  lengths  of  time.  The  effects  of  the 
maternal  diet  on  the  composition  of  the  unborn  young  were  not 
pronounced. 

Hoffstrom  (1910)  studied  metabolism  of  a  woman  during  preg- 
nancy, the  subject  receiving  an  ordinary  mixed  diet.  Among  the 
author's  conclusions  are  the  following: 

1.  The  phosphorus  excreted  through  the  urine  was  less  during 
the  second  half  of  the  period  than  the  first,  and  during  this  time  was 
comparatively  constant  in  amount. 

2.  The  organism  exhibits  a  greater  tendency  to  hold  back 
nitrogen  than  phosphorus. 

3.  The  average  phosphorus  storage  was  0.331  gm.  daily,  and 
the  cumulative  storage  of  phosphorus  reached  56  gm.  at  the  end  of 
the  experiment. 

4.  Of  this  amount  18  gm.  were  deflected  to  the  fetus,  and 
consequently  38  gm.  were  retained  by  the  mother  organism  itself. 
This  was  calculated  according  to  figures  of  Michel  for  phosphorus 
content  of  the  fetus  at  different  stages  of  development. 

Numerical  data  from  this  work  are  contained  in  the  following 
table : 


472 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


DAILY  PHOSPHORUS  METABOLISM  DURING  PREGNANCY 

Grama 


Week  of 

Total 

Cumula- 

Ratio 

Ratio 

Ratio 

preg- 

Intake 

Urine 

Feces 

outg-o 

Balance 

tive 

N:P 

N:P 

N:P 

nancy 

P 

P 

P 

P 

P 

retention 
P 

Intake 

Urine 

Feces 

17 

2.060 

0.955 

1.033 

1.988 

+0.072 

0.504 

5.8 

10.8 

0.9 

18 

2.300 

0.886 

0.823 

1.709 

+0.591 

4.641 

6.5 

12.7 

1.1 

19 

2.862 

1.178 

0.679 

1.857 

+1.005 

11.676 

6.1 

11.6 

1.2 

20 

2.471 

1.099 

1.195 

2.294, 

+0.177 

12.915 

6.4 

11.7 

1.0 

21 

2.304 

0.991 

1.000 

1.991 

+0.313 

15.106 

6.5 

12.4 

1.1 

22 

2.223 

1.006 

0.735 

1.741 

+0.482 

18.480 

6.5 

11.1 

1.6 

23 

2.231 

1.067 

0.781 

1.848 

+0.383 

21.161 

7.1 

10.7 

1.6 

24 

1.482 

0.774 

0.390 

1.164 

+0.318 

23.387 

7.0 

12.7 

1.4 

25 

1.927 

0.851 

0.854 

1.705 

+0.222" 

24.941 

7.1 

13.4 

1.0 

26 

1.835 

0.653 

0.784 

1.437 

+0.398 

27.727 

7.0 

15.4 

1.1 

27 

1.569 

0.763 

0.512 

1.275 

+0.294 

29.785 

7.0 

11.8 

1.2 

28 

1.458 

0.854 

0.536 

1.390 

+0.068 

30.261 

7.3 

11.7 

1.1 

29 

1.998 

0.907 

0.652 

1.559 

+0.439 

33.334 

6.9 

11.4 

1.3 

30 

1.754 

0.935 

0.694 

1.629 

+0.125 

34.209 

7.3 

10.4 

1.3 

31 

2.136 

0.966 

0.779 

1.745 

+0. 391 

36.946 

6.5 

9.8 

1.2 

32 

1.892 

0.895 

0.620 

1.515 

+0.377 

39.585 

7.1 

10.8 

1.2 

33 

1.834 

0.903 

0.677 

1.580 

+0.254 

41.363 

6.2 

10.5 

1.3 

34 

1.909 

0.845 

0.574 

1.419 

+0.490 

44.793 

6.3 

9.8 

1.5 

35 

1.994 

0.898 

0.523 

1.421 

+0.573 

48.804 

6.0 

9.2 

1.5 

36 

1.670 

0.919 

0.623 

1.542 

+0.128 

49.700 

6.7 

9.7 

1.6 

37 

1.957 

0.962 

0.607 

1.569 

+0.388 

52.416 

6.1 

9.2 

1.5 

38 

1.972 

0.936 

0.635 

1.571 

+0.401 

55.223 

6.4 

9.6 

1.9 

39 

1.872 

0.947 

0.710 

1.657 

+0.215 

56.728 

7.2 

11.5 

1.7 

40 

1.149 

0.832 

0.467 

1.299 

-0.150 

55.828 

6.2 

9.3 

1.3 

Mean 

1.952 

0.918 

0.703 

1.621 

+0.331 

6.6 

11.1 

1.3 

See  also  Mosler  (1853). 

The  extent  of  the  general  anabolic  excess  during  pregnancy  is 
indicated  by  the  work  of  E.  Landsberg  (1912),  who  found  in  human 
pregnancy  a  retention  of  nitrogen  four  times  as  great,  and  of 
sulphur  and  phosphorus  twice  as  great,  as  that  required  for  the 
fetus. 

E.  and  J.  Hermann  (1912)  reported  a  study  of  the  lipoid 
content  of  the  blood  of  normal  and  pregnant  women,  and  of  new-born 
infants.  The  phosphatid  content  is  found  about  the  same  for  the 
three  bloods,  though  the  proportion  of  the  total  fat  varies,  because 
both  cholesterin  esters  and  neutral  fat  are  high  in  the  blood  of  the 
pregnant  woman,  and  low  in  that  of  the  infant. 

SLEEP 

Laehr  (1890)  studied  metabolism  as  affected  by  sleep,  with 
himself  as  the  subject.  He  concluded  that  the  calcium,  magnesium 
and  phosphorus  outgo  were  not  influenced  by  sleep. 

Breisacher  (1891)  determined  phosphorus  and  nitrogen  in  the 
urine  in  three  8-hour  periods  per  day,  during  ten  consecutive  days, 
in  a  study  of  the  effects  of  sleep  on  metabolism.  No  figures  were 
given  on  the  composition  of  the  food.  Averages  of  the  daily  deter- 
minations are  as  follows : 

RELATION  OF  P20;  TO  N  IN  URINE 

12-8  A.  M P.03:N=1:5.29 

8  A.  M.-4  P.  M P205:N=1:7.46 

4  P.  M.-12 P205:N=1:6.93 


PHOSPHORUS  METABOLISM 


473 


The  subject  slept  during  the  period  from  12  P.  M.  to  8  A.  M. 
The  author  does  not  come  to  a  definite  conclusion  as  to  the  reason 
for  the  low  proportion  of  nitrogen  to  phosphorus  in  the  urine  formed 
during  sleep.  A  more  rapid  elimination  of  food  nitrogen  than  of 
food  phosphorus  would  produce  such  variation  in  the  rate  of  excre- 
tion as  the  author  noted. 

Sherman  (1902)  investigated  metabolism  as  affected  by  loss 
of  sleep  (see  table,  p.  432),  on  three  successive  nights.  Increased 
elimination  did  not  occur  until  the  third  day,  while  changes  resulting 
from  alteration  of  the  diet  were  always  perceptible  on  the  first  day. 
There  was  slight  increase  in  the  outgo  of  nitrogen,  sulphur  and 
phosphorus,  the  proportions  not  being  markedly  abnormal. 

PHOSPHORUS  METABOLISM  AS  AFFECTED  BY  THIRST  AND  WATER 

DRINKING 

Landauer  (1894)  states  that  deprivation  of  water  increases 
both  nitrogen  and  phosphorus  outgo,  but  that  after  reaching  a  cer- 
tain limit  the  phosphorus  elimination  returns  to  the  normal. 

Straub  (1899)  investigated  metabolism  during  thirst  in  dogs. 
The  phosphorus  elimination  in  the  urine  during  periods  of  3-4  days 
without  water  intake  was  undoubtedly  increased,  though  only  to  a 
slight  extent,  about  5-10  percent. 

In  connection  with  his  thorough  investigation  of  the  effects  of 
water-drinking  on  metabolism  Hawk  (1905)  has  studied  phosphorus 
metabolism,  with  results  as  indicated  below. 

DAILY  PHOSPHORUS  BALANCES  WITH  NORMAL  MATURE  MEN  ON  A 
STANDARD  DIET  WITH  VARYING  AMOUNTS  OF  WATER— Grams 


Subject 

Length 
of 

Initial 

Gain 

and 

period 

body 

Food 

Urine 

Feces 

or 

Diet 

period 

in 

days 

weight 
Kilos 

P2O5 

P2O5 

P2O5 

loss 
P2O5 

1-1 

2 

61.51 

5.226 

2.340 

2.39 

+0.496 

Milk  1800  g-m.,  butter  75  firm. 
330  g-m.,  water  500  c.  c 

crackers 

1-2 

2 

5.226 

2.650 

2.36 

+0.216 

Same  as  above  plus  4500  c  c.  water 

1-3 

4 

5.226 

2.310 

2.33 

+0.588 

Same  as  No.  1 

2-1 

1 

4.960 

2.509 

Milk  1650  g-m.,  butter  60  g-m., 
300  gm.,  water  900  c.  c. 

crackers 

2-2 

2.754 

Standard,  same  as  above 

2-3 

" 

2.406 

Standard 

2-4 

" 

2.568 

Standard  +  3100  c  C.  water 

2-5 

" 

2.653 

Standard  +  3100  c  c  water 

2-6 

** 

1.840 

Standard 

3-1 

" 

2.120 

" 

3-2 

" 

2.275 

'* 

3-3 

" 

2.330 

" 

3-4 

" 

2.479 

Standard  +  3100  c  c.  water 

3-5 

" 

2.700 

■  C                     •  <         <•             •(                 >• 

3-6 

** 

2.167 

Standard 

4-1 

2 

62.8 

•* 

2.925 

1.89 

+0.145 

*' 

4-2 

2 

" 

3.227 

2.20 

-0.462 

Standard  +  3100  c.  c.  water 

5-1 

2 

61.4 

" 

2.766 

1.92 

+0.279 

Standard 

5-2 

'  2 

" 

2.943 

2.02 

-0.003 

Standard  +  3100  c.  c.  water 

5-3 

2 

2.823 

1.93 

+0.207 

Standard 

474  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

From  this  work  the  author  concludes  that  copious  water  drink- 
ing increases  the  excretion  of  phosphorus  in  the  urine,  this  increase 
being  due  to  increased  cellular  activity,  and  the  accompanying  katab- 
olism  of  phosphorus-containing  bodies.  In  every  instance  the 
excretion  of  P205  was  increased  above  the  normal  on  each  day  of  the 
water  period,  the  maximum  excretion  occurring  on  the  second  day  of 
the  increased  ingestion. 

From  the  evidence  at  hand,  then,  one  must  conclude  that  either 
great  thirst  or  unusual  consumption  of  water  will  increase  the  phos- 
phorus excretion,  though  further  work  is  required  for  the  positive 
determination  of  the  source  of  this  phosphorus  and,  therefore,  the 
significance  of  this  increased  outgo. 

See  also  Breed  (1851a). 

EFFECTS  OF  TIME  OF  DAY  ON  PHOSPHORUS  METABOLISM 

This  is  one  of  those  subjects  which  seems  readily  explainable 
on  the  basis  of  a  few  results,  but  puzzling  after  the  accumulation  of 
an  abundance  of  evidence.  A  number  of  extended  studies  of  the 
question  have  been  made,  but  there  is  no  such  unanimity  of  findings, 
or  prospect  of  useful  conclusions,  as  inclines  one  to  attempt  to 
harmonize  and  summarize  the  results. 

From  the  papers  of  W.  Kaup  (1856b),  Haxthausen  (1860), 
Pettenkofer  and  Voit  (1866),  Edlefsen  (1878,  1881),  Speck  (1882), 
Breisacher  (1891),  Roeske  (1897),  Vogt  (1906)  and  Sarvonat  and 
Gentz  (1911)  we  conclude  that  this  is  a  matter  which  is  much 
influenced  by  personal  idiosyncrasy,  environment,  and  various  condi- 
tions of  the  general  method  of  life,  such  for  instance  as  the  time  of 
the  chief  meal  of  the  day  (W.  Zuelzer,  1876).  We  shall  mention  in 
detail  only  two  of  the  later  papers. 

Sherman  and  Hawk  (1900)  studied  the  urinary  elimination  by 
human  beings  of  nitrogen,  sulphur  and  phosphorus  as  affected  by 
the  time  of  day.  As  measured  by  3-hour  periods,  the  rates  of 
elimination  of  nitrogen  and  sulphur  run  closely  parallel,  and  normal- 
ly show  a  tendency  to  rise  during  the  morning,  reaching  a  maximum 
after  the  midday  meal,  with  a  slight  fall  in  the  following  period,  and 
another  rise  after  the  evening  meal.  During  the  night  the  excre- 
tion usually  reaches  the  minimum. 

The  excretion  of  phosphorus  describes  a  curve  altogether  differ- 
ent from  that  of  nitrogen  and  sulphates,  rising  steadily  from  the 
middle  of  the  morning  until  the  time  of  retiring,  then  falling  during 
the  hours  of  sleep,  and  continuing  to  fall  for  three  hours  after  rising, 
reaching  a  minimum  after  breakfast. 


PHOSPHORUS  METABOLISM 


475 


After  the  ingestion  of  a  considerable  amount  (63.7  gm.)  of 
extra  protein  .in  lean  beef  with  the  breakfast,  the  nitrogen  began  to 
rise  in  the  first  three  hours,  the  sulphur  and  phosphorus  a  little 
later.  The  nitrogen  excretion  regained  the  normal  in  36-39  hours, 
the  sulphur  a  little  earlier,  and  the  phosphorus  about  12-15  hours 
after  the  ingestion  of  the  beef. 

Hawk  and  Chamberlain  (1904)  also  studied  this  question  with 
results  as  below. 

VARIATION  IN  THE  COURSE  OF  PHOSPHATE  EXCRETION  IN  THE 

URINE  AS  AFFECTED  BY  TIME  OF  DAY  AND  INCREASED 

INGESTION  OF  PROTEIN 

Averages  of  Results  from  Two  Normal  Men — Grams 


July 

July 

July 

July 

July 

July  " 

July 

July 

July 

Period 

16 

17 

18 

19 

20 

21 

22 

23 

24 

1 
6:30  a.  m.— 9:30  a.  m. 

0.076 

0.086 

0.137 

0.127 

0.191 

0.165 

0.100 

•0.194 

0.149 

2 
9:30  a.  m— 12:30  p.  m. 

0.126 

0.191 

0.254 

0.257 

0.402 

0.317 

0.233 

0.325 

0.276 

3 
12:30  p.  m— 3:30  p.  m. 

0.302 

0.321 

0.411 

0.422 

0.509 

0.432 

0.383 

0.427 

0.409 

4 
3:30  p.  m. — 6:30  p.  m. 

0.229 

0.243 

0.308 

0.330 

0.409 

0.337 

0.383 

0.351 

0.349 

5 
6:30  p.  m— 9:30  p.  m. 

0.264 

0.329 

0.399 

0.339 

0.411 

0.390 

0.366 

0.431 

0.335 

6 
9:30  p.  m.— 6:30  a.  m. 

0.747 

0.882 

0.876 

0.881 

1.098 

1.007 

0.909 

0.850 

0.880 

7 
Total  for  day 

1.745 

2.050 

2.387 

2.358 

3.022 

2.702 

2.375 

2.580 

2.402 

Average  initial  weight  of  subjects,  56.14  kg.;  diet,  crackers  100  gm.,  butter  20  gm.,  whole 
milk  550  gm.  per  meal;  3  meals  per  day,  except  on  July  20,  when  the  morning  meal  was 
changed  to  crackers  50  gm.,  butter  20  gm.,  milk  450  gm.,  and  lean  beef  100  gm. 

In  the  excretion  of  phosphorus  in  the  urine  two  distinct  rises 
were  seen  each  day.  The  maximum  rate  was  reached  after  the 
midday  meal.  The  rate  of  excretion  reached  the  minimum  during 
the  first  period  in  the  morning.  The  maximum  rate  of  phosphate 
excretion,  due  to  the  ingestion  of  meat  in  the  morning  meal,  fell  in  a 
period  between  those  in  which  the  maxima  of  nitrogen  and  sulphate 
occurred,  that  is,  after  the  nitrogen  maximum  and  before  the 
sulphate  maximum.  With  one  subject  the  normal  rate  of  nitrogen 
and  phosphate  excretion  was  regained  on  the  second  day  following 
the  increased  proteid  ingestion ;  the  normal  rate  of  sulphate 
excretion  was  regained  one  day  later,  i.  e.,  three  days  after  the  inges- 
tion. With  the  other  subjects  the  normal  rate  of  excretion  was 
not  regained  in  any  case  until  the  fourth  day  following  the  increase 
in  proteid  food. 


476  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  6 

PART  V 
PHOSPHORUS  METABOLISM  IN  DISEASE 

ACID  INTOXICATION 

The  phenomenon  of  acidosis  is  a  general  condition  rather  than  a 
specific  disease,  and  is  a  symptom  in  many  disorders  of  nutrition. 
The  immediate  cause  is  a  disturbance  of  the  phosphate-carbonate 
balance  in  the  blood  which  constitutes  its  central  agency  for  the 
maintenance  of  that  approximate  neutrality  which  is  necessary  to 
the  function  of  respiration. 

Having  to  do  especially  with  compounds  other  than  those  of 
phosphorus,  we  shall  but  briefly  consider  this  important  matter,  and 
for  a  fuller  discussion  the  reader  is  referred  to  vonNoorden's  Metab- 
olism and  Practical  Medicine. 

The  general  subject  of  acid  intoxication  was  opened  up  by  the 
experiments  on  acid  poisoning  with  animals  by  F.  Walter  (1877). 
Acidosis  as  relating  to  human  nutrition,  however,  has  to  do  partic- 
ularly with  acid  products  of  disordered  metabolism,  rather  than  with 
acid  poisons  introduced  from  without,  and  this  field  of  learning  was 
developed  after  Stadelman  (1883)  discovered  /?-oxybutyric  acid,  and 
demonstrated  the  fact  that  diabetic  coma  is  at  least  largely  due  to 
this  compound. 

In  general,  excess  of  acid  in  the  system  may  be  brought  about 
in  two  ways,  first  by  actual  excess  of  acid  products,  and  second 
by  such  a  deficiency  of  bases  as  leaves  the  organism  subject  to 
disturbance  by  a  relative  excess  of  the  normal  acid  products  of 
katabolism,  the  one  being  in  effect  quite  as  truly  an  acid  excess  as 
the  other. 

The  acids  of  the  body  which  may  enter  into  the  disturbance  of 
its  approximate  neutrality  are  of  diverse  origin.  They  may  be  (1) 
inorganic  acids  from  outside  the  body,  (2)  unoxidized  organic 
products  of  acid  reaction,  (3)  acid  salts,  and  acid  ions  of  dissociated 
neutral  salts,  and  (4)  sulphuric  and  phosphoric  acids  resulting  from 
protein  katabolism ;  while  relative  excess  of  acids  may  come  about 

(1)  through  abnormally  low  intake  of  alkalis  and  alkaline  earths, 

(2)  through  excessive  outgo  of  these  same  elements,  and  (3) 
through  deficiency  of  intake  in  mineral  matter  generally,  for  even  a 
neutral  salt  may  serve  for  acid  neutralization  through  its  anion 
entering  into  relations  which  mask  its  potential  acidity,  as  for 
instance  with  proteins,  thus  freeing  its  cation. 


PHOSPHORUS  METABOLISM  477 

The  acids  actually  involved  in  the  production  of  acidosis  as 
we  know  it  in  practice  are  principally  /3-oxybutyric  and  related  com- 
pounds, in  actual  acid  excess,  and  the  sulphuric  and  phosphoric  acids 
of  normal  katabolism,  in  relative  excess  of  acids. 

The  reasons  are  somewhat  obscure  for  the  failure  of  the  body, 
in  acidosis,  to  oxidize  /?-oxybutyric  acid  in  the  normal  manner,  and 
for  its  consequent  appearance  along  with  the  derived  diacetic  acid 
and  acetone  in  the  urine.  They  appear,  however,  to  be  either 
deficiency  of  carbohydrates,  as  in  starvation  and  fever,  or  defective 
powers  of  oxidation  of  carbohydrates,  as  in  diabetes ;  the  coincident 
oxidation  of  carbohydrates  apparently  being  necessary  to  the  normal 
oxidation  of  /3-oxybutyric  as  well  as  other  fatty  acids. 

Actual  acidity  of  the  blood  in  the  living  animal,  however,  is 
impossible.  These  acids  do  not  circulate  free,  but  are  neutralized 
for  the  protection  of  the  organism.  Normally  the  alkalis  used  for 
the  neutralization  of  the  acid  products  of  katabolism  are  furnished 
principally  (1)  by  the  oxidation  of  organic  salts  of  the  fixed  alkalis 
and  alkaline  earths  in  the  food  (especially  among  herbivora) ,  (2)  by 
the  separation  of  acid  urine  from  alkaline  blood,  (3)  by  metabolic 
ammonia  deflected  from  urea  formation  (especially  among  omnivora 
and  carnivora) ,  (4)  also  to  a  less  extent  by  carbonates  and  ammonia 
in  the  food,  and  (5)  by  ammonia  split  off  from  proteins  in  the 
alimentary  tract  by  bacteria. 

The  automatically  variable  agency  for  the  compensation  of 
varying  amounts  of  acid  in  the  system  is  the  above-mentioned  deflec- 
tion, for  acid  neutralization,  of  a  part  of  the  ammonia  normally 
contributed  by  the  tissues  to  the  blood  as  carbamate,  and  synthe- 
sized to  urea  in  the  liver,  the  acids  thus  forming  ammonium  salts, 
and  appearing  as  such  in  the  urine.  The  amount  of  ammonia  thus 
rendered  available  for  acid  neutralization  is  quite  considerable 
(omnivora  and  carnivora) ,  but  at  the  same  time  it  is  not  unlimited, 
and  the  protection  afforded  is  not  perfect,  even  as  far  as  it  goes, 
since  the  ammonium  salts  themselves  become  somewhat  toxic 
through  ionization  (Wilbur,  1904) . 

Our  usual  measure  of  acid  excess  in  the  system,  then,  however 
produced,  and  of  whatever  nature,  is  the  ammonia  of  the  urine  (that 
is,  total  ammonia — free  NH3  and  NH3  in  salts),  though  an  actual 
estimation  of  the  disturbing  compounds  in  the  blood,  as  well  as  the 
estimation  of  the  inorganic  elements  of  the  excreta,  afford  us  supple- 
mentary information. 


478  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

In  brief,  the  effects  of  acidosis  are  dyspnoea,  and  attendant 
nervous  symptoms,  terminating  in  stupor  and  collapse,  the  causes 
being  the  expulsion  of  carbon  dioxide  from  the  blood  by  stronger 
acids,  its  accumulation  in  the  tissues,  and  the  resultant  interference 
with  their  oxygenation. 

Herbivora  and  carnivora  differ  greatly,  as  Walter  showed,  as  to 
their  resourcefulness  in  adjusting  themselves  to  excess  of  acids.  In 
herbivora  the  food  normally  furnishes  a  superabundance  of  alkali, 
and  therefore  these  animals  have  neither  need  nor  capacity  for  the 
extensive  use  of  ammonia  in  acid-neutralization  which  is  character- 
istic of  flesh-eating  animals.  Acid  poisoning,  therefore,  causes  a 
much  greater  increase  in  the  excretion  of  fixed  bases  by  herbivora 
than  by  omnivora  and  carnivora. 

So  far  as  known  the  prevalence  of  acidosis  as  differentiated 
from  calcium  starvation  in  domestic  animals  has  not  yet  been 
demonstrated. 

In  human  beings  acidosis  is  well  known  in  a  great  variety  of 
diseases  and  conditions,  especially  in  diabetes,  gastrointestinal  dis- 
orders in  children,  fever,  starvation,  pregnancy,  burns  and  anaes- 
thesia. 

Taylor  (1904)  made  a  study  of  human  metabolism  on  an  ash- 
free  diet.  Metabolism  was  disturbed  apparently  by  sulphuric  and 
phosphoric  acids  of  metabolic  origin. 

VonNoorden  (1907)  states  that  the  blood,  muscles  and  glands 
give  up  no  appreciable  amounts  of  alkali  in  diabetic  acidosis,  but 
that  there  is,  at  the  same  time,  a  loss  from  the  body,  apparently 
from  the  bones,  of  calcium,  magnesium  and  phosphorus.  By  subse- 
quent work  of  many  other  investigators  these  facts  are  well  estab- 
lished. 

In  gastrointestinal  disorders  of  children  there  is  an  acidosis 
which  was  explained  by  the  work  of  Keller,  Steinitz  and  Freund. 
Keller  (1897)  found  an  increased  elimination  of  ammonia  in  the 
urine  always  accompanying  an  increase  in  the  fat  of  the  food.  Stein- 
itz (1903)  showed  that  this  increased  ammonia  excretion  was  due  to 
.an  acidosis  caused  by  a  withdrawal  of  alkali  from  the  body  in  soaps 
formed  from  fats  in  the  intestine,  and  excreted  in  the  feces.  W. 
Freund's  (1905)  observations  make  it  apparent  that  the  increased 
soaps  in  the  feces  are  calcium  soaps,  a  part  of  the  calcium  normally 
excreted  as  calcium  phosphate  uniting  with  the  fatty  acids  to  form 
soaps,  leaving  an  equivalent  amount  of  the  phosphorus  to  unite  with 
alkalis  and  ammonia,  the  urinary  excretion  of  which  is  thereby 
increased. 


PHOSPHORUS  METABOLISM  479 

VonJaksch  (1885)  first  showed  that  the  acidosis  of  fever  is  of 
the  acetone  variety.  The  acetone  bodies,  /?-oxybutyric  acid,  diacetic 
acid  and  acetone,  are  much  less  abundant  in  fever  than  in  diabetes, 
and  also  less  abundant  than  in  complete  inanition. 

In  general,  considering  the  causes  and  degrees  of  acidosis  in 
fevers,  the  quantitative  excretion  of  the  acetone  bodies  is  controlled 
much  more  directly  by  inanition,  fat  destruction,  the  seat  of  the 
infection,  and  the  individuality  of  the  patient  than  by  the  temper- 
ature, or  by  the  severity  of  the  infection. 

As  bearing  on  the  effects  of  experimentally  produced  dyspnoea 
on  phosphorus  elimination,  Saccone  (1907)  found  that  restriction  of 
respiration  in  a  dog,  by  the  application  of  a  Sayre  corset,  caused  an 
increase  in  urinary  phosphorus  which  disappeared  after  two  days. 

We  may  consider,  then,  that  the  connection  of  acid  intoxication 
with  phosphorus  metabolism  is  through  the  participation  of  phos- 
phates in  the  maintenance  of  neutrality ;  through  the  contribution  of 
phosphoric  acid  to  the  total  acids  of  the  body ;  through  the  interfer- 
ence of  abnormal  amounts  of  acid  with  synthesis  and  retention  of 
phosphorus  compounds,  especially  in  the  bones;  and  otherwise  in 
still  more  general  ways. 

A  few  references  to  articles  which  we  have  consulted  on  acid 
intoxication  are  the  following:  Allard  (1907) ;  Ewing  (1908) ;  Folin 
(1907) ;  Nichols  (1908) ;  Rossi,  F.,  :  Inner  Respiration  of  Tissues 
and  Its  Relation  to  Death  from  Hunger  and  to  Acidosis  (Bui.  Sci. 
Med.,  81, 149-54) ;  Szili  (1906) ;  Talbot  (1907)  and  Fitz,  Alsberg  and 
Henderson  (1907). 

ACROMEGALY 

That  this  disease  is  due  to  an  excess  of  function  of  the  hypoph- 
ysis, or  pituitary  body,  is  the  prevailing  opinion,  and  one  of  the 
more  satisfactory  investigations  in  support  of  this  hypothesis  is  the 
work  of  Lewis  (Johns  Hopkins  Hosp.  Bui.  16  (1905),  157.),  who 
found,  by  histological  demonstration,  hyperplasia  of  the  chromophile 
cells  of  the  anterior  lobe,  and  other  conditions  indicative  of  glandular 
activity. 

Schiff  (1897a,  1897b)  used  hypophysis  and  thyroid  preparations 
in  metabolism  studies  with  acromegalics.  The  nitrogen  and  phos- 
phorus of  the  diets  were  estimated  by  calculation,  but  in  the  urine 
and  feces  were  determined  by  analysis.  He  found  that  both  the 
hypophysis  and  thyroid  preparations  react  strongly  on  the  sluggish 


480  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

metabolism  of  acromegalics,  producing  diuresis,  and  increased  nitro- 
gen and  phosphorus  excretion,  with  protracted  after-effects,  and 
loss  of  weight. 

Tauszk  and  Vas  (1899)  noted  retention  of  phosphorus  and  loss 
of  calcium  in  acromegaly,  and  negative  results  from  the  use  of 
pituitary  and  thyroid  tablets. 

VonMoraczewski  (1901)  conducted  metabolism  studies  in  acro- 
megaly, with  analyses  of  food,  urine  and  feces,  and  with  blood  exam- 
ination. Hypophysis  and  thyroid  tablets  produced  no  change  in  the 
phosphorus  retention,  which  was  considerable  throughout  this 
experiment.  In  a  second  experiment  treatment  with  yohimbine, 
elemental  phosphorus,  silver  nitrate  and  oxygen  all  lessened  the 
retention,  or  caused  a  loss  of  phosphorus. 

Oswald  (1902),  experimenting  with  a  dog,  found  no  definite 
peculiarities  of  metabolism  following  ingestion  of  powdered  pituitary 
body  in  quantities  of  1-2  gm. 

Edsall  and  Miller  (1903)  conducted  a  well-controlled  metabo- 
lism study  of  acromegaly.  The  percentage  of  retention  of  phos- 
phorus was  20.88,  nitrogen  12.86,  and  calcium  9.32  in  a  7-day  period. 
The  phosphorus  retention  was  over  a  gram  a  day.  The  relation  of 
stored  calcium  and  phosphorus  shows  that  the  latter  could  not  have 
been  used  to  any  considerable  extent  in  the  growth  of  bone.  The 
subject  was  an  imbecile. 

Malcolm  (1904)  studied  metabolism  in  the  dog  as  affected  by 
the  ingestion  of  pituitary  gland  substance.  The  glandular  and 
"nervous"  portions  of  the  gland  were  fed  separately  in  different 
periods,  and  in  others  the  whole  gland.  The  calcium  and  phosphorus 
balances  led  the  author  to  the  conclusion  that  the  nervous  portion  is 
probably  the  more  active  and  has  the  effect  to  cause  the  katabolism 
of  bone  tissue. 

Franchini  (1904)  published  results  of  an  experiment  in  the 
feeding  of  hypophysis  tablets  to  a  case  of  acromegaly.  This  treat- 
ment produced  nothing  decisive  in  results  on  phosphorus  metabo- 
lism. 

In  a  later  study  (1908b)  Franchini  published  blood  analyses 
from  three  cases  of  acromegaly.  The  data,  however,  are  insufficient 
to  warrant  the  drawing  of  conclusions.  Still  later  (1910)  Franchini 
published  metabolism  and  blood  data  on  rabbits  and  guinea  pigs 
under  injection  treatment  with  hypophysis  extracts  prepared  from 
cattle  and  horses.  These  extracts  caused  a  marked  loss  of  calcium 
and  magnesium  and  a  less  marked  loss  of  phosphorus.  In  the  circu- 
lating blood  was  found  an  increase  of  calcium  and  magnesium. 


PHOSPHORUS  METABOLISM  481 

Thompson  and  Johnston  (1905)  made  metabolism  studies  on 
dogs  under  pituitary  treatment  by  ingestion  per  os.  The  feces  were 
not  analyzed  for  phosphorus.  Pituitary  substance  appears  to  stim- 
ulate metabolism  in  the  dog,  as  shown  by  increased  outgo  of  nitrogen 
(urine  and  feces),  of  urea,  and  to  a  less  degree  of  urinary  phos- 
phates, effects  which  do  not  pass  away  immediately  when  the  feed- 
ing of  pituitary  body  is  discontinued. 

Diesing  (1909)  expresses  the  opinion  that  the  hypophysis  gland 
controls  phosphorus  metabolism,  and  that  its  function  is  physiologic- 
ally opposite  to  that  of  the  thyroid  in  such  sense  that  when  the 
hypophysis  stores  too  much  phosphorus,  too  little  connective  tissue 
is  formed,  leading  to  myxoedema,  or  in  younger  individuals  to 
cretinism,  having  the  same  effect  as  atrophy  of  the  thyroid.  When 
the  hypophysis  is  below  normal  it  causes  an  excessive  growth  of 
bone  and  connective  tissue.      The  disease  acromegaly  is  the  result. 

Medigreceanu  and  Kristeller  (1911)  injected  sterile  extract  of 
anterior  lobes  of  hypophysis  into  an  acromegaly  patient  on  two  days 
during  a  20-day  period  of  observation.  Previous  to  the  first  injec- 
tion the  patient  was  in  phosphorus  equilibrium.  The  injection 
caused  a  marked  loss  of  phosphorus.  Between  the  first  and  second 
injections  the  organism  showed  a  decided  tendency  to  reestablish 
equilibrium,  but  on  the  second  injection  a  marked  loss  of  phospho- 
rus was  again  produced. 

See  also  Delille,  (1909). 

Mochi  (1909)  studied  elimination  of  nitrogen,  calcium,  magne- 
sium and  phosphorus  in  the  urine  of  8  starving  rabbits,  of  which  6 
were  treated  by  injection  with  extract  of  fresh  lamb  hypophysis,  one 
with  extract  of  nerve  substance,  and  one,  the  control,  left  without 
treatment.  The  author  found  that,  as  compared  with  the  control, 
the  animals  treated  with  extract  of  hypophysis  died  sooner  than  the 
others,  but  lost  less  in  weight;  further,  the  relation  between  the 
nitrogen  and  phosphorus  of  the  urine  showed  such  an  excess  of  the 
latter  as  to  indicate  destruction  of  bone  tissue.  The  relation  of 
nitrogen  to  P205  in  the  urine  was  as  follows :  Control,  6.5:1 ;  treated 
with  hypophysis,  4.6,  4.3,  4.5,  5.9,  5.3  and  4.5 :  1,  and  treated  with 
extract  of  nerve  substance  8.1:1.  (Bibliography  of  26  refer- 
ences.) 

Mochi  (1910)  reported  further  experiments  with  extract  of 
pituitary  body  injected  into  fasting  guinea  pigs.  In  complete  balance 
experiments  he  learned  that  the  increased  loss  of  phosphorus  under 
the  pituitary  treatment  is  without  doubt  due  to  tearing  down  of 
bone  tissue.     A  part  of  the  numerical  results  are  as  follows : 


482 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


AVERAGE  DAILY  LOSSES  OF  NITROGEN,  PHOSPHORUS  AND  CALCIUM 
BY  GUINEA  PIGS  TREATED  WITH  EXTRACT  OF  HYPOPHYSIS 

Grams 


No,  of 

Treatment 

Duration 

of 

experiment 

in  days 

N 

P 

Ca 

Partition  of  loss 
of  phosphorus 

animal 

From 
muscle 

From 
hone 

I 

n 

in 

IV 

Fasting1  only 
Fasting  and  injection 

fC                   cc                   «< 

18 
17 
16 
15 

1.6436 
0.3718 
0.4376 
1,0127 

0.0934 
0.1516 
0.1069 
0.1050 

0.0943 
0.2776 
0.1571 
0.1154 

0.0183 
0.0215 
0.0498 

0.1334 

0.0854 
0.0552 

Rubinraut  (1912)  studied  metabolism  in  acromegaly,  and  deter- 
mined that  hypophysis  ingestion  increased  calcium  and  phosphorus 
elimination,  and  caused  marked  loss  of  these  elements,  while  thyroid 
therapeutics  decreased  the  rate  of  elimination  of  calcium  and  phos- 
phorus in  proportion  to  the  amount  of  thyroid  preparation  ingested,, 
the  balances  becoming  positive.  Potassium  iodide  resembled  hypoph- 
ysis in  its  effects,  increasing  to  a  considerable  extent  both  calcium 
and  phosphorus  outgo. 

Aschner  (1912)  has  made  a  recent  and  extensive  study  of  the 
function  of  the  hypophysis.  By  extirpation  experiments  with  dogs 
he  obtained  results  varying  in  intensity  inversely  as  the  age  of  the 
animal.  In  very  young  dogs  the  removal  of  the  hypophysis  had 
the  effect  to  retard  general  development  in  marked  and  characteris- 
tic ways,  affecting  the  size,  hair,  claws,  teeth,  bones,  sexual  develop- 
ment, etc.  These  effects  were  produced  by  removal  of  the  whole 
gland,  or  the  frontal  lobe,  not  by  removal  of  the  posterior  lobe. 

Aschner  notes  the  contradictory  evidence  on  results  of  hypoph- 
ysis administration,  and  cites  a  case  of  his  own,  a  hypoplastic 
dwarf  in  whom  there  was  supposed  to  be  under-functioning  of  the 
hypophysis,  who  under  hypophysis  treatment  increased  in  height  7 
cm.  in  5  months. 

Aschner  cites  the  work  of  Benda,  M.  Sternberg,  Erdheim,  B. 
Fischer,  v.  Frankl-Hochwart,  Marburg  and  others  in  support  of  his 
belief  that  acromegaly  is  caused  by  over-functioning  or  disordered 
functioning  of  the  hypophysis,  a  part  of  the  symptoms  being  due  to 
its  internal  secretion,  and  a  part  to  its  local  action  on  surrounding 
parts  of  the  brain.  The  true  dwarf  condition  appears  to  be  due,  at 
least  largely,  to  hypo-functioning  of  the  hypophysis,  while  giantism 
is  rather  definitely  associated  with  an  excessive  functioning  of  this 
gland. 

The  connection,  therefore,  of  this  matter  with  phosphorus 
metabolism  is  general  in  character,  but  with  an  especial  bearing  on 
the  growth  of  the  bones. 


PHOSPHORUS  METABOLISM  483 

ALCOHOLISM 

Romeyn  (1887)  showed  that  the  ingestion  of  35-50  c.c.  of 
alcohol  by  fasting  men  temporarily  decreased  urinary  phosphorus 
elimination,  an  effect  which  was  followed  in  a  few  hours,  first  by  an 
increased  elimination,  and  then  soon  by  a  decrease  toward  the 
normal  for  the  fasting  subject.  The  increased  nitrogen  elimination 
was  much  less  marked. 

Saccone  (1907)  found  that  alcoholic  intoxication  reduced  the 
urinary  phosphorus  in  dogs,  due  perhaps  to  retarded  metabolism. 

Salant  and  Hinkel  (1910),  experimenting  with  4  dogs,  found 
that  subacute  alcohol  intoxication  in  well-fed  dogs  causes  a  moder- 
ate diminution  of  total  nitrogen  and  total  sulphur,  and  a  much 
greater  diminution  of  total  and  inorganic  sulphates  and  of  phos- 
phates in  the  urine.  There  is  also  a  tendency  to  retention  of  chlor- 
ides. 

Schittenhelm  (1909a)  notes  that  both  Pollak  and  Bloch  recog- 
nize the  fact  that  in  alcoholism,  as  in  gout,  there  is  characteristic 
delay  and  diminution  of  uric  acid  elimination,  and  that  Landau  in 
extensive  studies  has  come  to  the  conclusion  that  the  elimination  of 
endogenous  purins  is  increased,  but  that  exogenous  uric  acid  is 
decreased  by  the  loss  of  ability  of  the  kidneys  to  let  uric  acid  pass, 
a  phenomenon  as  yet  unexplained. 

Schittenhelm  conducted  experiments  with  dogs  in  an  effort  to 
clear  up  this  point.  Since  dogs  excrete  their  purins  mostly  as  allan- 
toin,  while  men  excrete  the  same  as  uric  acid,  it  was  thought  that  if 
there  were  disturbance  in  ferment  action,  which  results  in  slowing 
up  excretion,  it  would  probably  show  as  a  change  in  the  end 
products;  and  if  on  the  other  hand  there  is  a  specific  injury  to  the 
kidney  filtration  of  uric  acid,  it  would  hardly  hold  to  the  same  extent 
for  the  more  easily  soluble  allantoin. 

Chronic  alcoholism  was  established  by  feeding  meat  scrap 
treated  with  ethyl  and  amyl  alcohol;  and  daily  urinary  nitrogen 
studies  were  made  both  during  feeding  and  fast.  As  in  alcohol  experi- 
ments with  men  there  was  delayed  purin  elimination,  as  shown,  in 
this  case,  by  the  allantoin.  The  dog  had  no  appearance  of  nephri- 
tis, and  microscopic  examination  of  the  kidneys  showed  no 
evidence  of  disease,  from  which  it  would  appear  that  there  was 
delayed  purin  metabolism. 

Baskoff  (1909b)  found  that  alcohol  poisoning  reduced  the  leci- 
thin content  of  the  liver  of  the  dog. 

Choumova-Sieber  (1910;  same  as  Sieber,  1909a)  studied  the 
effects  of  alcoholism  on  the  organism  by  the  administration  of  daily 
doses  of  alcohol  to  three  dogs  during  several  months.      One  or  two 


484  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

months  after  the  last  dose  they  were  killed,  and  the  organs  analyzed 
for  phosphatids.  Compared  with  three  control  animals  they  showed 
a  decrease  in  the  phosphatid  content  of  all  the  organs  examined 
except  the  kidneys:  brain  11.41  percent,  mucosa  of  stomach,  4.15 
percent,  intestinal  mucous  membrane  3.22  percent,  spleen  3.18  per- 
cent, liver  2.74  percent,  heart  2.52  percent,  lungs  1.34  percent,  intes- 
tinal wall  1.10  percent,  and  others.  The  percentage  of  decrease 
was  reckoned,  not  on  the  phosphatid  content  of  the  organs,  but  on 
the  total  dry  matter  of  these  parts,  and  hence  the  losses  noted  must 
be  considered  as  astonishingly  large.  Examination  of  the  individ- 
ual records  in  both  groups  shows  that  in  almost  all  cases  each  of  the 
controls  exceeds  all  of  the  experimental  subjects  in  the  phosphatid 
content  of  the  organs.  This  article  contains  a  bibliography  of  56 
references. 

PHOSPHORUS  METABOLISM  IN  ARTHRITIS  AND  CHRONIC 
RHEUMATISM 

Stokvis  (1876),  studying  phosphorus  excretion  in  arthritis, 
finds  that  the  urinary  phosphorus  combined  with  calcium  and  mag- 
nesium is  considerably  reduced  during  the  whole  of  the  attack,  and 
that  even  after  the  ingestion  of  lime  or  magnesia  the  earthy  phos- 
phates remain  much  reduced  in  the  urine,  while  in  a  normal  subject 
increased  intake  of  calcium  carbonate  or  magnesia  increases  the 
earthy  phosphates  of  the  urine. 

Godefroy  (1903)  finds  in  the  urine,  in  chronic  rheumatism,  sub- 
normal amounts  both  of  phosphates  and  total  phosphorus,  but  on 
the  other  hand  much  more  than  the  normal  amount  of  incompletely 
oxidized  phosphorus. 

Joulie  (1904)  administered  phosphoric  acid  to  rheumatic  horses 
in  doses  of  10  c.c.  of  the  acid  (sp.  gr.  1.35)  diluted  to  one  liter.  The 
excretion  of  calcium  phosphate  by  the  kidneys  is  said  to  have  been 
increased. 

See  also  vonNoorden  and  Belgard  (Berliner  klin.  Wochenschr., 
1894). 

BERIBERI 

Beriberi  is  a  nutritional  disorder  especially  prevalent  among 
the  rice-eating  peoples  of  the  world,  but  is  not  unknown  even  in 
America,  where  it  has  been  found  in  certain  fishing  settlements  of 
Labrador  in  which  the  people,  during  several  months  of  the  year, 
live  largely  on  white  bread  and  tea.  The  disease  is  characterized 
by  profound  nervous  disorganization,  and  appears  to  be  essentially 
the  same  as  the  multiple  polyneuritis  of  fowls. 

Volumes  have  been  written  to  show  that  beriberi  is  caused  by 
deficiency  of  phosphorus  in  the  food ;  also  great  works  to  show  that 


PHOSPHORUS  METABOLISM  485 

it  is  due  to  poisons  of  fungous  origin.  Both  of  these  positions  have 
been  generally  abandoned,  and  the  problem  seems  to  be  at  least  near 
to  solution,  if  indeed  it  is  not  already  solved,  on  quite  another  basis. 
It  is,  therefore,  considered  unnecessary  to  review  the  earlier  litera- 
ture of  the  subject. 

Prevailing  opinion  as  to  the  dietetic  basis  for  the  disease  was 
set  forth  by  a  resolution  of  the  Far  Eastern  Association  of  Tropical 
Medicine  after  a  conference  at  Manila,  in  March  1910.  The  resolu- 
tion was  as  follows : 

"Resolved,  that  in  the  opinion  of  this  Association  sufficient 
evidence  has  now  been  produced  in  support  of  the  view  that  beriberi 
is  associated  with  the  continuous  consumption  of  white  (polished) 
rice  as  the  staple  article  of  diet,  and  the  Association  accordingly 
desires  to  bring  this  matter  to  the  notice  of  the  various  govern- 
ments concerned." 

H.  Schaumann  (1910)  has  given  us  an  exhaustive  treatise  on 
beriberi  in  relation  to  the  phosphorus  of  the  food,  and  for  a  review 
of  the  literature  up  to  1910  the  reader  is  referred  to  this  mono- 
graph. 

His  position  at  the  time  of  this  publication  was  that  beriberi 
is  a  metabolism  disease  which  is  due  to  an  insufficient  supply  of 
organic  phosphorus  to  the  tissues,  this  being  caused,  in  general,  by 
too  small  a  content  of  organic  phosphorus  in  the  food,  and  in  other 
cases,  apparently  much  less  frequently,  by  insufficient  absorption  of 
organic  phosphorus  compounds  when  they  are  supplied  in  sufficient 
quantities  in  the  food. 

Schaumann  considers  that  scurvy,  Barlow's  disease,  rickets  and 
osteomalacia  may  have  a  similar  etiology.  As  indicating  some  of 
the  details  of  opinion  at  that  time,  and  subsequently,  we  submit 
the  following  notes. 

Fraser  and  Stanton  (1910)  conclude  that  beriberi  is  caused 
by  white  rice,  and  that  the  tendency  of  rice  to  cause  beriberi  varies 
inversely  as  the  total  phosphorus. 

De  Haan  (1910)  coincides  with  these  conclusions,  and  finds  that 
neither  lack  of  salts  nor  nucleins  in  the  rice  is  the  cause  of  beriberi. 

Highet  (1910)  agrees  in  his  findings  with  the  above. 

Shibayama  (1910)  considered  monotony  of  diet  as  a  predispos- 
ing factor.  He  believed  the  disease  to  be  due  to  a  specific  organ- 
ism. 

Kilbourne  (1910)  reported  that  beriberi  varied  in  prevalence 
in  companies  of  native  Philippine  scouts  inversely  as  the  phosphorus 
and  potassium  of  the  diet. 


486 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


Aron  (1910a)  reported  that  diets  such  as  cause  beriberi  contain 
insufficient  phosphorus  to  maintain  health,  and  that  diets  similar 
to  those  thought  to  cause  beriberi  cause  polyneuritis  in  chickens. 
Experiments  with  chickens  showed  that  the  addition  of  phytin  was 
of  some  benefit  in  this  condition. 

Aron  and  Hocson  (1910)  reported  the  following  balance  data: 

AVERAGE  DAILY  PHOSPHORUS  AND  NITROGEN  BALANCES 
Periods  Four  Days  Each — Grams 


Period 

and 
subject 

P2O5 

N 

Food 

Intake 

Urine 

Feces 

Out- 
put 

Bal- 
ance 

Bal- 
ance 

a 

1-normal 

Polished    rice,    bread, 
sugar,  dried  fish,  coffee 

1.674 

1.138 

0.526 

1.664 

+0.01 

+0.38 

Rice  diet 

b 
1-normal 

Same  4-  75    gm.    rice 
bran,  (less  bread  and 
rice;  more  sugar  than 
above) 

5.460 

1.378 

3.460 

4.838 

+0.62 

+0.57 

Phosphorus  added 
as  rice  bran 

c 

2-normal 

Polished    rice,    bread, 
sugar,  coffee,  bacon 

1.498 

1.735 

0.733 

2.468 

-0.97 

-4.67 

Lower  in  N  and  P 
than  above 

d 
2-normal 

Same  +   12  gm.    egg 
albumen,  (less   sugar, 
morecoffee  than  above) 

1,878 

1.568 

0.610 

2.178 

-0.30 

-1.98 

Protein    added    to 
above 

e 

3-beriberi 
patient 

Same  as  above  +  6  gm. 
phytin,     (more    coffee 
than  above) 

4.913 

1.563 

1.845 

3.408 

+1.50 

-2.08 

Phosphorus,  as 
phytin,  ,  added   to 
above 

f 

3-beriberi 

patient 

Rice,     bread,      sugar, 
coffee,  bacon 

1.103 

1.553 

0.730 

2.283 

—1.18 

-2.29 

Diet  such  as  sup- 
posed     to      cause 
beriberi 

g 

3-beriberi 

patient 

Same    as    above    but 
with  bacon  containing 
less  N  and  P2O5 

1.045 

1.240 

0.843 

2.083 

-1.03 

-1.94 

Diet  such  as  sup- 
posed     to      cause 
beriberi 

h 
3-beriberi 
patient 

Same  as  above  +  6  gm. 
phytin 

3.700 

1.313 

2.885 

4.198 

-0.50 

-1.81 

Phosphorus,  as 
phytin,  added  to 
above 

l 
3-beriberi 
patient 

Same    as    above    but 
with  less    bread    and 
coffee 

3.638 

1.093 

3.090 

4.183 

-0.54 

-2.77 

Phosphorus,  as 
phytin,  added  to 
above 

i 

3-beriberi 
patient 

Same    as    above  _  but 
with  more  and  differ- 
ent bacon 

1.013 

0.703 

0.655 

1.358 

—0.35 

-1.22 

No  phosphorus 
added 

k 
3-beriberi 
patient 

Same  as   above  +  12 
gm.  egg  albumen,  but 
with  less  sugar 

1.263 

0.840 

0.790 

1.630 

-0.37 

—2.09 

Protein  added 

1 

3-beriberi 
patient 

Rice,     bread,     sugar, 
fish 

1.908 

1.133 

0.913 

2.045 

-0.13 

-0.58 

Typical     Filipino 
diet;  about  same  as 
first  ration  above 

From  this  work  some  of  their  conclusions  are  as  follows : 
"A  diet  consisting  of  bread  and  rice  (both  poor  in  phosphorus), 
some  fat  (bacon)  and  sugar,  furnishing  40  calories,  0.15  gm.  N  and 
0.025  gm.  P2Or,  per  kilo  body  weight  does  not  cover  the  demands  of 
the  body  for  N  and  P205,  and  therefore  leads  to  N  and  P205  loss 
from  the  body.  Addition  of  protein  reduces  the  N  loss  of  the  body 
and  the  loss  of  P205. 


PHOSPHORUS  METABOLISM  487 

"The  addition  of  phosphorus  in  the  form  of  phytin  prevents  a 
loss  of  that  constituent  from  the  body,  and  if  sufficient  of  this 
element  is  added  a  storage  of  phosphorus  after  a  period  of  phospho- 
rus starvation  takes  place.  The  loss  of  nitrogen  from  the  body 
is  reduced  by  the  addition  of  phytin,  as  compared  with  a  corre- 
sponding period  in  which  phytin  is  not  given. 

"A  diet  consisting  of  fish,  bread,  rice,  sugar,  etc.,  furnishing  37 
calories,  0.2  gm.  N  and  0.032  gm.  P205  per  kilo  of  body  weight,  is 
sufficient  to  keep  a  man  in  N  and  P205  equilibrium. 

"The  addition  of  rice  bran  has  a  tendency  to  produce  a  slight 
storage  of  P205;  the  rice  polish  in  this  respect  corresponding  to 
phytin.  The  phosphorus,  both  of  rice  and  of  phytin,  is  excreted 
almost  entirely  in  the  feces. 

"It  is  highly  probable  that  living  for  an  extended  period  on  a 
one-sided  almost  exclusively  vegetable  diet,  which  is  characterized 
by  its  poverty  in  phosphorus  and  in  protein,  may  result  in  beriberi. 

"The  process  of  polishing  rice  removes  a  fine  skin  and  the  outer 
layers  (bran) ;  this  rice  bran  is  rich  in  phosphorus,  especially  in  its 
organic  soluble  form  (phytin) ;  the  content  of  phosphorus  of  the 
rice  is  considerably  reduced  by  the  removal  of  the  bran. 

"Polished  rice,  poor  in  phosphorus,  may  cause  beriberi  in  man 
if  it  is  the  main  constituent  of  the  food,  but  it  is  harmless  if 
sufficient  other  nourishment,  rich  in  phosphorus  and  protein,  is 
taken.  The  same  polished  rice  causes  a  polyneuritis  in  chickens. 
White  bread,  a  food  of  similar  chemical  composition  as  regards 
phosphorus  and  protein,  cannot  sustain  monkeys  in  normal  health 
if  it  forms  the  entire  diet. 

"The  addition  of  phytin  (the  organic  phosphorus  compound  of 
rice  bran)  considerably  reduces  the  deleterious  effects  of  white  rice 
on  chickens." 

In  connection  with  their  study  of  rice  as  the  cause  of  beriberi 
Aron  and  Hocson  (1911a,  1911b)  report  the  following  balance  data 
(see  next  page)  from  experiments  with  healthy  men. 

The  subjects  subsisted  on  diets  composed  largely  of  rice  of 
different  conditions  as  to  milling.  The  husked  rice  contained  0.7- 
0.8  percent  P205,  undermilled  rice  0.45-0.60  percent  P205,  and  over- 
milled  rice  0.15-0.35  percent  P205.  In  addition  to  rice  the  subjects 
received  bread,  fish,  sugar,  bacon,  coffee,  etc.  The  results  show  that 
an  intake  of  less  than  1.65  gm.  P205  per  50  kg.  of  body  weight 
(0.033  gm.  P205  per  kg.)  is  insufficient  to  cover  the  demands  of  the 
body  for  phosphorus.      The  only  cases,  Nos.  3,  6,  and  11,  in  which 


488 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


the  balance  was  positive  were  those  which  received  unpolished  rice, 
rice  bran  or  phytin.  Experiments  Nos.  3  and  4  show  that  the  body 
loses  phosphorus  on  a  diet  of  white  rice,  and  that  when  this  is 
replaced  by  red  rice,  the  amount  of  phosphorus  exceeds  the  demand 
of  the  body.  When  the  phosphorus  intake  was  high  the  excess 
was  excreted  almost  entirely  in  the  feces. 

AVERAGE    DAILY    PHOSPHORUS    METABOLISM    WITH    HUMAN 
SUBJECTS  ON  DIETS  COMPOSED  PRINCIPALLY  OF  DIFFERENT 
SORTS  OF  RICE— Grams 


No.  and 
length  of 

Subject 

Body 
weight 
Kilos 

Food 
P2O5 

Outgro  P2O5 

Balance 
P2O5 

P2O5  metabolism 

per  50  kg-,  body 

weight 

experiment 

Urine 

Feces 

Intake 

Balance 

3 
4  days 

Prisoner  C 

49.0 

3.22 

1.03 

2.03 

+0.16 

3.30 

+0.16 

4 
4  days 

.. 

48.6 

1.42 

1.04 

0.83 

—0.45 

1.45 

-0.45 

5 
4  days 

B 

64.0 

1.50 

1.74 

0.73 

-0.97 

1.15 

-0.80 

6 
4  days 

" 

64.0 

4.91 

1.56 

1.85 

+1.50 

3.85 

+1.20 

7 
4  days 

.. 

64.0 

1.88 

1.57 

0.61 

-0.30 

1.45 

-0.30 

9 
6  days 

D 

43.5 

1.28 

0.80 

0.49 

-0.01 

1.65 

-0.01 

10 
4  days 

A 

52.5 

1.57 

1.14 

0.52 

+0.01* 

1.60 

+0.01* 

11 
4  days 

.. 

52.5 

5.46 

1.38 

3.46 

+0.62 

5.20 

+0.60 

12 
3  days 

E 

54.0 

1.77 

1.23 

0.76 

-0.22 

1.65 

-0.20 

13 
3  days 

.. 

54.0 

1.77 

1.11 

0.73 

-0.10 

1.65 

-0.10 

14 
3  days 

G 

45.9 

'  1.45 

0.95 

0.58 

-0.08 

1.60 

-0.10 

*  To    harmonize   with    other    data,    should    he   — 0.09. 

Jebbink  (1910)  concluded,  from  dietetic  studies,  that  deficiency 
of  nuclein  phosphorus  in  the  ration  is  responsible  for  the  production 
of  beriberi. 

Janin  (1910)  got  sufficient  benefit  from  the  administration  of 
organic  phosphorus  compounds  to  lead  him  to  believe  that  deficiency 
of  these  in  the  diet  causes  beriberi. 

Kajiura  and  Rosenheim  (1910),  as  also  Eijkmann,  found  that 
barley,  fed  with  rice,  relieves  symptoms  of  polyneuritis  in  poultry, 
and  that  this  improvement  is  not  due  to  the  alcohol-soluble  proteid. 


PHOSPHORUS  METABOLISM  489 

Fujitani  (1910)  found  that  the  very  thin  skin  of  half-hulled 
rice  has  the  power  to  protect  from  polyneuritis,  but  although  its 
phosphorus  was  present  mostly  as  phytin,  that  phytin  itself  does 
not  produce  the  same  benefit. 

Chamberlain,  Bloombergh  and  Kilbourne  (1910)  found  that 
potassium  and  phosphorus  salts  did  not  cure  polyneuritis,  and  that 
the  idea  of  deficiency  of  phosphorus  as  the  cause  of  beriberi  must 
be  abandoned.  These  authors  found  polyneuritis  curable  by  a 
dialyzable  substance  which  could  be  extracted  from  rice  polish  with 
either  cold  water  or  cold  alcohol. 

Teruuchi  (1910)  found  that  phosphorus  excretion  is  not  very 
different  in  beriberi  from  the  same  under  normal  conditions; 
further,  that  the  protective  principle  in  unhusked  rice  and  in  oats, 
while  not  injured  by  100°  heat,  is  destroyed  by  heating  to  130°-135°. 
Teruuchi  confirmed  the  conclusions  of  Eijkmann,  Fraser  and  Stanton 
that  the  neuritis-protecting  principle  in  rice  hulls  is  soluble  in  warm 
alcohol,  and  that  it  is  probably  not  a  phosphorus  compound. 

Simpson  and  Edie  (1911)  report  that  an  exclusive  diet  of  white- 
flour  bread,  with  pigeons,  produced  degenerative  changes  in  the 
peripheral  nerves,  and  death;  while  a  diet  of  whole-wheat  bread 
resulted  in  health  and  gain  in  weight.  These  authors  cured  this 
disease  by  the  feeding  of  yeast. 

Chamberlain  and  Vedder  (1911)  report  the  multiple  polyneu- 
ritis of  poultry  and  beriberi  as  essentially  the  same.  They  found 
that  extracts  of  rice  polish,  and  also  of  white  beans,  would  cure 
polyneuritis.  Chamberlain  (1911)  also  notes  the  disappearance  of 
beriberi  from  the  native  Philippine  scouts  with  a  change  of  diet. 

Breaudat  and  Denier  (1911)  also  cured  beriberi  with  rice 
bran. 

Fraser  and  Stanton  (1911a,  1911b)  note  the  fact  of  the  low 
phosphorus  content  of  rice  which  causes  beriberi,  and  present  a  gen- 
eral review  of  the  evidence  as  to  the  cause  of  beriberi.  They  state 
that  cooking  of  the  unpolished  rice  under  pressure  at  120°  for  two 
hours  destroys  that  principle  which  prevents  polyneuritis.  Attempts 
to  isolate  this  active  principle  are  described.  The  protective  sub- 
stance was  found  to  be  soluble  in  0.3  percent  HC1,  and  in  alcohol. 
Phytin  was  shown  not  to  be  the  protective  compound. 

Cooper  and  Funk  (1911)  report  that  exclusive  diets  of  starch, 
inulin,  cane  sugar,  and  dextrine  will  cause  polyneuritis  in  poultry; 
that  phytin,  edestin,  casein,  or  egg-yolk  will  not  cure  the  same, 
but  that  yeast  both  prevents  and  cures  polyneuritis.  Yeast  press 
juice  also  cures,  as  does  yeast  juice  after  24  hours  hydrolysis  with 


490  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

20  percent  sulphuric  acid.  Another  curative  preparation  was  made 
from  rice  polish  by  solution  in  alcohol,  precipitation  by  phosphotung- 
stic  acid,  and  decomposition  with  baryta.  This  compound  was 
free  from  phosphorus,  carbohydrates  and  proteins. 

Schaumann  (1911a)  concludes  that  the  protective  effect  of 
various  preparations  such  as  phytin,  yeast-lecithin,  pepsin-hydro- 
chloric acid  extract  of  phaseolus,  rice-bran-phosphatid,  and  purified 
hydrochloric  acid  extract  of  rice  bran  and  yeast,  is  due  to  the  com- 
bined presence  of  organic  phosphorus  and  an  "activator,"  which 
acts  on  these  compounds  in  such  manner  as  to  render  them  com- 
pletely available  to  the  organism. 

Chamberlain  and  Vedder  (1912)  concluded  that  beriberi  was 
not  due  to  phosphorus  shortage  nor  to  acid  intoxication. 

Funk  (1912a)  found  that  polyneuritis  of  birds  causes  a 
decreased  nitrogen  and  phosphorus  content  of  the  brain,  and  is 
caused  by  the  lack  of  a  basic  alcohol-soluble  substance  which  is 
necessary  to  the  functioning  of  the  nervous  tissues ;  that  the  medul- 
lary sheath,  both  within  and  without  the  central  nervous  system,  is 
more  or  less  degenerated,  that  polyneuritis  can  be  cured  by  prepara- 
tions from  milk,  yeast,  and  lime  juice;  that  the  cure  is  very  rapid, 
the  functioning  of  the  axis  cylinder  being  restored  before  the  regen- 
eration of  the  medullary  sheath  is  completed. 

Edie,  Evans,  Moore,  Simpson  and  Webster  (1912)  isolated  from 
yeast  an  anti-neuritic  basic  compound  which  they  call  "torulin,"  and 
with  which  they  plan  to  conduct  experiments  relating  to  beriberi. 
The  approximate  percentage  composition  was  C  40.5,  H  8.07,  N 
13.32,  O  38.11.  Three  possible  structural  formulae  are  suggested 
as  follows: 

C7H17N205 

C7H16N02(HN03) 

N(CH3)3C4H702(HN03) 

Suzuki,  Shamimura  and  Odake  (1912)  have  isolated  a  basic 
compound  from  rice  bran  which  they  believe  to  be  the  active  prin- 
ciple in  the  prevention  of  beriberi.  It  is  an  organic  compound, 
soluble  in  alcohol,  insoluble  in  ether,  is  precipitated  by  phosphotung- 
stic  acid  from  solutions  acidified  by  sulphuric  acid,  and  from  water 
solution  by  tannin.  It  was  isolated  in  crystalline  form  as  the 
picrate.  This  compound  they  call  "orizanine."  Of  the  pure  com- 
pound obtained  from  the  picrate,  5-10  mg.  given  per  os  or  subcutane- 
ously  suffice  to  cure  a  pigeon  suffering  from  rice  polyneuritis.  Simi- 
lar results  were  obtained  with  chickens,  mice  and  dogs.  They 
regard  orizanine  as  an  essential  nutrient.      Dogs  fed  on  meat  and 


PHOSPHORUS  METABOLISM  491 

polished  rice  die  in  a  few  weeks  with  symptoms  of  starvation.  They 
remain  in  health  if  0.3  gm.  of  orizanine  is  added  to  the  daily  diet. 
Judged  from  their  therapeutic  action,  grains  and  vegetables  in  gen- 
eral contain  orizanine.  Milk,  eggs,  fish  and  meat,  and  their  alco- 
holic extracts  were  without  effect  on  pigeons  suffering  from  the 
polished-rice  disease.  With  dogs,  however,  the  alcoholic  extract 
of  meat  is  practically  as  effective  as  orizanine  in  preventing  or 
curing  the  disease.  Details  of  the  chemistry  of  the  compound  are 
given. 

Schaumann  (1912a)  now  practically  accepts  the  position  and 
conclusions  of  Funk  and  his  associates,  and  expresses  his  belief, 
based  on  experimental  observations,  that  the  anti-neuritic  extract 
with  which  he  is  working  contains  a  phosphatese,  or  several  such, 
and  suggests  that  this  "activator"  serves  alike  in  plant  and  animal 
physiology  in  playing  a  part  in  the  synthesis  of  organic  phosphorus 
compounds. 

Wieland  (1912)  fed  mice  on  polished  rice,  and  studied  the 
organic  and  inorganic  phosphorus  of  their  bodies  with  reference  to 
Schaumann's  earlier  hypothesis  connecting  beriberi  with  insuffi- 
ciency of  certain  organic  phosphorus  compounds  in  the  diet.  His 
results  did  not  sustain  the  theory. 

MacLean  (1912b)  found  a  substance  in  animal  tissues  which 
possesses  an  anti-neuritic  capacity.  It  is  soluble  in  water  and  in 
alcohol.  It  is  not  a  lipoid,  but  is  precipitated  by  excess  of  acetone 
from  both  its  water  and  alcohol  solutions.  It  contains  phosphorus 
apparently  only  as  an  impurity.  This  substance  is  likely  to  be  pres- 
ent as  an  impurity  in  lecithin  as  ordinarily  prepared,  and  MacLean 
suggests  that  to  this  fact  must  be  due  the  variable  results  from  the 
use  of  lecithin  in  beriberi. 

Eijkmann  (1911)  found  that  the  protective  principle  in  rice 
hulls  is  effective  after  peritoneal  injection. 

Fraser  and  Stanton  (1914)  state  that  a  satisfactory  measure 
of  the  degree  of  polishing  of  rice,  in  relation  to  the  cause  of  beriberi, 
is  its  total  phosphorus  content.  A  rice  which  contains  less  than 
0.4  percent  P2Og  cannot  safely  be  permitted  to  form  the  staple 
article  of  a  diet  for  man. 

See  also  Moszkowski  (1911),  Vedder  and  Clark  (1912),  Vedder 
(1912),  Strong  and  Crowell  (1912),  Gouzien  (1912),  Fargier  (1912), 
Hulshoff-Pol  (1912),  Heiser  (1912),  Grijns  (1912),  Yamigawa, 
Koyana,  Midorikawa  and  Mogi  (1912),  Onodera,  Nakamura  and 
Tateno  (1912)  and  Gregg  (1913). 


492       OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

THE  BLOOD  IN  RELATION  TO  PATHOLOGICAL  PHOSPHORUS 

METABOLISM 

A  review  of  the  literature  of  the  diseases  of  the  blood,  and  of 
the  composition  of  the  blood  in  disease  reveals  a  considerable 
measure  of  uncertainty  and  disagreement  in  the  fragmentary 
evidence,  and  comparatively  little  which  is  characteristic  in  the 
associated  phosphorus  metabolism,  a  condition  due  in  part,  at  least, 
to  the  natural  variability  in  the  composition  of  the  blood  in 
harmony  with  the  number  and  diversity  of  its  general  service  func- 
tions. 

The  bearing  of  the  blood  diseases  on  nuclein  metabolism  as 
indicated  by  uric  acid  excretion  has  been  discussed  somewhat 
recently  by  H.  Strauss  in  von  Noorden's  Metabolism  and  Practical 
Medicine,  to  which  we  refer  the  reader.  We  shall,  however,  review 
a  portion  of  the  literature  of  other  aspects  of  the  subject. 

PHOSPHORUS  METABOLISM  IN  CHLOROSIS  AND  ANAEMIA 

Vannini  (1904),  in  five  balance  experiments,  in  a  study  of 
chlorosis,  found  absorption  of  fats,  carbohydrates  and  proteins 
normal,  and  a  nitrogen  storage  in  four  cases  out  of  five.  The  obser- 
vations on  the  mineral  nutrients  seem  to  us  neither  significant  nor 
characteristic.  With  calcium,  magnesium  and  phosphorus  the 
positive  balances  were  about  as  numerous  as  the  negative  balances, 
but  the  latter  slightly  exceeded  the  former  in  magnitude. 

VonMoraczewski  (1897c)  also  conducted  balance  experiments 
with  3  cases  of  chlorosis,  and  2  of  anaemia  from  carcinoma.  In 
anaemia  there  was  retention  of  nitrogen,  calcium  and  phosphorus 
in  both  of  the  cases.  In  chlorosis  there  was,  in  each  case,  nitrogen 
retention,  but  the  results  with  calcium  and  phosphorus  varied  with 
the  haemoglobin  content  of  the  blood.  With  25  percent  of  the 
normal  haemoglobin  there  was  loss  of  both  calcium  and  phosphorus; 
with  40  percent  haemoglobin  there  was  retention  of  calcium  and  loss 
of  phosphorus ;  with  60  percent  haemoglobin  there  was  retention  of 
both  calcium  and  phosphorus.  The  addition  of  sodium  chloride  and 
CaHP04  to  the  food  caused  nitrogen  economy.  VonMoraczewski 
concluded  that  anaemia  itself  causes  no  protein  decomposition. 

Stejskal  and  Erben  (1900b)  report  results  of  a  balance  experi- 
ment on  a  fatal  case  of  pernicious  anaemia.  Absorption  from  the 
alimentary  tract  was  poor.  The  salts  in  the  urine  were  related, 
one  to  another,  quantitatively,  as  in  the  food;  and  as  to  calcium, 
magnesium  and  phosphorus  the  patient  was  practically  in  equilibri- 
um.    There  was  a  slight  storage  of  nitrogen.     The  authors  stated 


PHOSPHORUS  METABOLISM  493 

the  opinion  that  pernicious  anaemia  is  caused  by  inanition  or  auto- 
intoxication following  stomach-intestinal  atrophy,  or  to  a  disturb- 
ance of  function  of  the  intestinal  mucous  coat. 

Von  Moraczewski  (1900b)  published  results  of  a  study  of  four 
cases  of  pernicious  anaemia.  He  states  that  the  results  show,  first 
of  all,  the  impaired  power  of  assimilation  of  the  organism.  There 
is  but  little  loss  of  nitrogen  on  nitrogen-poor  food,  and  little  storage 
of  nitrogen  on  adding  nitrogen  to  the  food.  Oxidation  appeared 
deficient,  as  in  chlorosis,  leukaemia  and  nephritis.  The  increased 
calcium  outgo  indicates  katabolism  of  bone  substance.  The  addi- 
tion of  lime  caused  a  general  improvement  of  symptoms,  with 
decreased  nitrogen  outgo,  and  improved  oxidation  of  sulphur  com- 
pounds. The  main  characteristics  of  pernicious  anaemia  he  states 
as  follows : 

(1)  Increased  lime  excretion;  (2)  the  small  values  of  the  out- 
go, especially  of  nitrogen  and  phosphorus ;  (3)  apathy  of  the  organ- 
ism; (4)  lack  of  accommodation  of  the  outgo  to  the  intake;  and  (5) 
usually  increased  (relatively)  chlorine  excretion. 

PHOSPHORUS  METABOLISM  IN  LEUKAEMIA 

Von  Moraczewski  (1898b)  published  balance  experiments  with 
one  case  each  of  leukaemia  and  pseudoleukaemia.  The  following 
are  among  his  conclusions : 

Leukaemia  is  a  nitrogen  and  phosphorus  disease ;  pseudoleukae- 
mia is  a  nitrogen  disorder.  Almost  all  constituents  of  the  food  were 
found  to  be  retained  abnormally  in  leukaemia,  because  of  a  lack  of 
katabolism,  more  in  respect  to  phosphorus  and  nitrogen  than  to 
chlorine  and  calcium.  In  the  case  of  pseudoleukaemia  the  nitro- 
gen and  calcium  elimination  was  impaired  to  a  greater  extent  than 
that  of  the  chlorine  and  phosphorus.  With  the  case  of  leukaemia 
the  inhalation  of  oxygen  caused  an  increased  calcium  and  phospho- 
rus outgo.  The  addition  of  sodium  chlorate  and  calcium  phosphate 
caused  an  increase  in  the  nitrogen  and  phosphorus  outgo,  and  an 
increase  in  the  red  blood  corpuscles,  with  a  decrease  of  leucocytes. 
The  treatment  with  thyroid  tablets  was  beneficial ;  elimination  was 
increased  and  the  balances  became  normal.  The  retention  of  phos- 
phorus is  noted  by  the  author  as  being  contrary  to  the  theory  that 
leucocytosis  should  cause  phosphorus  excretion. 

Milroy  and  Malcolm  (1898)  observed  that  in  splenomedullary 
leucocythaemia  the  pathological  condition  seemed  to  consist  in  a 
hindrance  to  the  leucocytic  breaking-down,  as  evidenced  by  an  abso- 
lute diminution  of  phosphorus,  and  a  marked  relative  decrease  of 


494  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

the  same  in  proportion  to  the  total  nitrogen  in  the  urine.  The 
alloxuric  nitrogen,  as  a  whole,  was,  relatively  to  the  total  nitrogen, 
increased  in  leucocythaemia,  both  the  bases  and  the  uric  acid  appar- 
ently participating  in  this  increase. 

In  a  later  publication  Milroy  and  Malcolm  (1899)  presented 
conclusions  from  a  further  study  of  leucocythaemia.  In  a  case  of 
lymphatic  leucocythaemia  they  found  a  marked  diminution  in  the 
absolute  amount  of  phosphorus  excreted,  and  also  a  relative  decrease 
compared  to  the  amount  of  nitrogen  excreted.  The  alloxuric  excre- 
tion (uric  acid  and  bases)  was  hardly  affected. 

In  a  case  of  medullary  leucocythaemia  where  the  number  of 
leucocytes  was  rapidly  falling,  the  phosphorus  excretion  was  found 
to  have  undergone  no  diminution,  while  the  alloxuric  excretion 
underwent  marked  diminution ,  which  the  authors  suggest  might  be 
explained  by  Ehrlich's  theory  of  leucocythaemia  being  a  form  of 
active  leucocytosis,  due  to  the  action  of  some  body  circulating  in  the 
blood,  and  acting  as  a  positive  chemotactic  agent  on  the  myelocytes 
and  fully  formed  polymorphonuclear  leucocytes  in  the  marrow. 

White  and  Hopkins  (1899)  published  urine  analyses  for  two 
leukaemic  patients,  in  comparison  with  those  from  two  normal  sub- 
jects. The  feces  were  not  analyzed.  They  observe,  in  harmony 
with  von  Moraczewski  (1898b),  the  lack  of  relation  between 
leucocytosis  and  the  excretion  of  products  such  as  phosphorus  and 
alloxuric  bodies,  which  result  from  katabolism  of  nucleins. 

Stejskal  and  Erben  (1900a)  conducted  complete  balance  experi- 
ments with  cases  of  lymphatic  and  myelogenic  leukaemia.  In  one 
very  anaemic  case  of  chronic  lymphatic  leukaemia,  on  a  normal 
mixed  diet,  there  was  in  a  5-day  experiment  a  marked  nitrogen 
retention,  5.2  gm.  per  day,  indicating,  according  to  the  authors, 
diminished  powers  of  oxidation.  Coincident  with  this  nitrogen 
storage  was  a  retention  of  0.17  gm.  P205  and  0.8  gm.  chlorine,  and  a 
marked  loss,  0.35  gm.,  of  calcium.  Of  magnesium,  the  output 
exactly  equalled  the  intake.  The  authors  attribute  the  loss  of 
calcium  to  decomposition  of  bone. 

In  a  case  of  leukaemia  Uenalis  et  myelogenes,  with  the  blood  in 
much  better  condition  than  in  the  above  case,  and  with  a  greater 
intake  of  nitrogen,  there  was  a  slight  loss  of  this  element  as  well 
as  of  phosphorus,  while  there  were  slight  gains  of  calcium  and  mag- 
nesium. 

According  to  Balthazard  (1901b)  an  increased  lecithin  content 
of  the  liver  is  general  in  diseases  involving  the  destruction  of 
leucocytes. 


PHOSPHORUS  METABOLISM  495 

Y.  Henderson  and  Edwards  (1903)  studied  one  case  of  lym- 
phatic leukaemia  during  a  period  of  six  and  a  half  months.  In 
spite  of  an  enormous  leucocytosis  (175000-380000  per  cu.  mm.,  of 
which  96  percent  were  lymphocytes),  and  in  spite  of  the  alternation 
of  periods  of  great  increase  and  marked  diminution  in  the  number 
of  circulating  corpuscles,  the  excretion  of  phosphorus  and  of  uric 
acid  was  at  no  time  excessive.  The  leucocytosis  was  considered  as 
due,  not  to  a  general  increase  in  nuclein  metabolism,  but  to  a  failure 
in  the  normal  destructive  processes.  During  the  progress  of  this 
study,  the  phosphorus  elimination,  which  was  subnormal  at  the 
beginning,  decreased  to  a  much  smaller  figure.  The  nitrogen,  also 
subnormal  at  the  beginning,  also  decreased,  but  not  to  so  great  an 
extent  as  the  phosphorus ;  so  that  there  was  an  increase  in  the  pro- 
portion of  nitrogen  to  phosphorus  in  the  urine.  At  the  same  time 
the  uric  acid,  which  was  somewhat  above  normal  at  the  beginning, 
increased  its  proportion  to  total  nitrogen  to  a  considerable  extent, 
and  its  proportion  to  phosphorus  very  greatly,  while  the  absolute 
uric  acid  excretion  increased  slightly.  The  decreased  nitrogen  and 
phosphorus  outgo  were  sjiown  to  be  due,  not  to  decreased  nuclein 
katabolism,  for  uric  acid  excretion  increased  during  the  study,  but 
to  retention. 

Henderson  and  Edwards  harmonize  the  conflicting  observations 
of  Milroy  and  Malcolm  (1898,  1899),  who  noted  decreased  phospho- 
rus excretion,  and  of  von  Moraczewski  (1898b),  who  noted  a  marked 
phosphorus  retention,  of  Magnus-Levy  (1898),  who  recorded  a 
great  increase  in  phosphorus  excretion  during  40  hours,  with 
slight  leucocytosis,  and  of  White  and  Hopkins,  who  concluded  that 
there  is  no  necessary  proportionality  between  the  number  of  circu- 
lating leucocytes  and  the  excretion  of  those  products  (P2Og  and 
alloxuric  bodies)  which  result  from  the  breakdown  of  nucleins,  by 
showing  that  these  apparent  differences  are  probably  due  to  these 
experimenters  having  made  their  observations  at  different  stages 
in  the  progress  of  the  disease,  as  made  clear  by  the  changes  in 
amount  and  proportion  of  urinary  constituents  during  their  own 
much  more  protracted  observations. 

Symmers  (1904-5a)  found  the  excretion  of  organic  phosphorus 
in  the  urine  pronounced  in  lymphatic  leukaemia,  there  being  a 
marked  change  in  the  ratios  of  organic  phosphorus,  both  to  total 
phosphorus  and  to  nitrogen,  while  the  ratios  of  nitrogen  to  inorgan- 
ic and  total  phosphorus  showed  little  if  any  departure  from  the 
normal;  and  Erben  (1908)  found  in  leukaemia  that  the  plasma  was 
normal,  but  the  erythrocytes  were  low  in  iron  and  potassium,  and 
high  in  sodium,  chlorine,  total  phosphorus  and  lecithin. 


496  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

PHOSPHORUS  METABOLISM  AS  AFFECTED  BY  BLOOD  DISSOLUTION 

Kiihnau  (1896-7)  experimented  on  a  considerable  number  of 
dogs  by  subcutaneous  and  intraperitoneal  injections  of  various 
poisons  and  foreign  substances  as  follows :  pyrogallic  acid,  blood  of 
a  dog  poisoned  by  pyrogallic  acid,  blood  of  a  normal  dog,  red  blood 
corpuscles  from  normal  dog  blood  suspended  in  physiological  salt 
solution,  magnesium  sulphate  plasma  of  a  pyrogallic-acid-poisoned 
dog,  serum  of  a  pyrogallic-acid-poisoned  dog,  normal  dog's  blood  ser- 
um, normal  human  blood  serum,  typhoid  serum,  leukaemia  serum, 
NaCl  solution  of  haemoglobin,  sodium  indoxylsulphate,  cinnabar  and 
pure  carbon.  Among  the  author's  conclusions  are  the  following:  In 
blood  dissolution  there  occur  considerable  leucocytosis,  and  the  fol- 
lowing changes  in  metabolism;  (a)  an  increased  excretion  of  uric 
acid  and  purin  bases,  (b)  a  rise  at  the  beginning,  in  the  excretion  of 
phosphoric  acid,  with  considerable  diminution  following  it,  (c)  an 
increased  excretion  of  chlorine,  and  (d)  chlorine  and  phosphoric  acid 
excretion  stand  in  an  inverse  relation  to  each  other,  in  that  increased 
excretion  of  the  one  corresponds  to  retention  of  the  other. 

PHOSPHORUS  OF  THE  BLOOD  AS  AFFECTED  BY  BLOOD  DISEASES 

Anaemia.  Von  Moraczewski  (1896)  observed  that  in  lead  colic 
and  anaemia  the  blood  relations  are  identical,  and  the  urine  also 
shows  the  same  similarity.  In  these  cases  there  was  an  increase 
of  chlorine  and  decrease  of  phosphorus  in  the  blood,  and  in  the 
urine  a  decrease  of  chlorine  and  increase  of  phosphorus,  especially 
the  alkaline  earth  phosphates. 

Masing  (1911a)  made  a  comparison  of  the  blood  of  rabbits  and 
geese,  in  a  normal  state,  with  the  same  after  the  production  of  an 
anaemic  condition  by  bleeding,  and  by  injection  of  phenylhydrazine. 
In  rabbit  serum  both  the  total  and  lipoid  phosphorus  were  increased 
by  the  production  of  an  anaemic  condition,  thus  differing  from 
chlorosis,  as  observed  by  Erben  (1902) ;  and  in  the  erythrocytes  the 
proportions  of  total,  lipoid  and  nuclein  phosphorus  to  total  nitrogen 
were  increased  by  the  production  of  anaemia.  In  the  goose  blood 
similar  results  were  obtained.  The  increase  in  the  lipoid  and 
nuclein  phosphorus  of  the  corpuscles  is  regarded  as  evidence  of  the 
youth  of  these  cells. 

Chlorosis.  Erben  (1902)  reported  analyses  of  the  blood  of 
three  chlorotics.  He  found  the  lecithin  content  of  the  serum  low, 
as  also  the  phosphorus  of  the  ash  of  the  serum,  pointing  to  a  decom- 
position of  erythrocytes.  There  was  an  increase  above  normal  in 
the  calcium  and  magnesium  of  the  serum.      In  the  erythrocytes 


PHOSPHORUS  METABOLISM  497 

the  fat  and  lecithin,  as  well  as  the  ash  constituents,  were  above 
normal;  the  cholesterin  subnormal;  while  the  iron  is  decreased  in 
proportion  to  haemoglobin.  Considering  the  blood  as  a  whole,  the 
protein,  lecithin,  cholesterin,  phosphorus,  potassium  and  iron  were 
subnormal,  while  the  fat,  calcium  and  magnesium  were  above  nor- 
mal. 

Leukaemia.  E.  Freund  and  Obermayer  (1891)  report  an  anal- 
ysis of  leukaemic  blood,  and  compare  it  with  analyses  of  normal 
blood  by  Jarisch,  Schmidt,  and  Becquerel  and  Rodier.  A  part  of 
the  figures  are  as  follows: 

COMPARISON  OF  COMPOSITION  OF  ASH  OF  LEUKAEMIC  AND 
NORMAL  BLOOD 


Leukaemic 

Normal 

(Freund  and  Obermayer) 

(Jarisch) 

P205 

16.92 

8.82 

so3 

12.31 

7.11 

CI 

17.82 

30.74 

K20 

15.65 

26.55 

Na20 

38.52 

24.11 

CaO 

0.47 

0.90 

MgO 

0.07 

0.53 

Fe303 

2.24 

8.16 

104.00 

106.92 

0  equivalent 

to  CI 

4.00 

6.92 

100.00  100.00 

Erben  (1908)  also  reports  a  case  of  leukaemia  in  which  the 
lecithin  and  total  phosphorus  of  the  erythrocytes  were  in  excess  of 
the  normal. 

PHOSPHORUS   OP  THE  BLOOD  AS   AFFECTED  BY  VARIOUS   DISEASES 

Erben  reported  analyses  of  the  blood  and  also  of  the  serum  and 
erythrocytes  in  a  number  of  pathological  conditions.  Data  from 
his  articles  of  1902,  1903,  1905,  1907  and  1908  are  included  in  the 
table  on  the  following  page. 

Von  Moraczewski  (1896)  published  extensive  series  of  blood 
and  urine  analyses  of  patients  with  pneumonia,  nephritis,  syphilis 
and  other  diseases.  In  one  case  of  pneumonia  there  was  in  the 
blood  a  subnormal  chlorine  content,  followed  by  an  increase  after 
the  crisis,  and  a  high  phosphorus  content  due  to  leucocytosis.  See 
table  on  page  499. 

Peritz  (1908)  studied  the  relation  of  lecithin  to  lues,  tabes  and 
paralysis.  In  the  first  contribution,  figures  were  reported  for  the 
ether-soluble  phosphorus  in  the  feces  of  cases  of  tabes  and  tabo- 
paralysis  as  affected  by  injections  of  lecithin.  The  results  are  of 
doubtful  significance. 


498 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


HUMAN  BLOOD  AS  AFFECTED  BY  VARIOUS  DISEASES 


Diagnosis 

Parts 

per  1000  of  substance 

Percent  of  ash 

Portion  of 
blood 

Leci- 
thin 
Grams 

P2O5 
Grams 

CaO 

Grams 

Mg-O 
Grams 

P2O5 

CaO 

Mg-O 

2.193 
1.769 
1.715 
1.668 
3.545 
1.455 
2.421 
1.226 

1.452 
0.964 
5.800 

1.536 
1.601 
5.232 
2.251 
2.387 
1.060 

1.822 
0.805 

2.391 

2.97 

2.15 

0.957 

3.163 

2.000 
1.780 
1.817 

0.292 
0.206 
0.228 

0.476 
0.477 
0.341 
0.120 

0.247 

0.764 

0.093 
0.144 
0.100 
0.130 
0.219 
0.095 

0.277 
0.097 

0.460 

0.57 

0.53 

0.873 

0.127 

0.111 

0.135 
0.257 
0.238 

0.219 
0.180 
0.175 
0.117 

0.145 

0.165 

0.355 
0.247 
0.357 
0.359 
0.232 
0.156 

0.177 
0.185 

0.067 

0.01 

0.21 

0.085 

0.202 

0.164 

0.077 
0.049 
0.055 

0.033 
0.043 
0.025 
0.012 

0.024 

0.060 

0.054 
0.065 
0.008 
0.005 
0.006 
0.009 

0.056 
0.077 

0.113 

0.03 

0.02 

0.063 

0.042 

0.036 

2.46 
2.33 
8.82 
5.83 
6.11 
4.12 
7.31 

3.17 

8.64 

1.14 
1.63 
1.26 
1.52 
2.68 
1.18 

3.26 
1.18 

3.08 
2.43 
0.90 
2.68 
2.30 
2.11 
1.58 

1.86 

1.87 

4.09 
2.79 
4.48 
4.21 
2.83 
1.94 

2.02 
2.25 

0,59 

ii        ii 

0.56 

•  I        it 

0.53 

::    :: 

Nephritis,  subchronic  parenchymatous 
Nephritis,  chronic  parenchymatous. .... 
Same  with  secondary  shrunken  kidneys 

Nephritis,    chronic    parenchymatous,  * 
shrunken  kidneys,  uremia,  hypertrophy 

0.40 
0.55 
0.30 
0.16 

0  31 

•i    ii 

•1    ii 

0.68 

O.fift 

0,74 

••    •• 

Nephritis,  subchronic  parenchymatous 

Nephritis,  chronic  parenchymatous 

Same  with  secondary  shrunken  kidneys 

Chronic    uremia,   secondary  shrunken 

0.10 
0.06 
0.07 
0.11 

0.66 

NephritiSi  chronic  parenchymatous*. . . . 

0.08 

■  • 

<  < 

*  Also  shrunken  kidneys,  uremia,   and  hypertrophy  of  the  heart. 


In  another  article  (1908-9a)  Peritz  states  that  he  has  found 
that  the  feces  of  patients  having  tabes  or  tabo-paralysis  were  rich 
in  lecithin,  while  in  the  normal  man  the  feces  contain  only  a  trace 
or  0.2  to  0.7  gm.  per  day.  There  was  also  an  increase  in  the  lecithin 
of  the  serum.  The  lecithin  content  of  the  serum  in  tabes  and  par- 
alysis is  given  as  4.7  to  6.15  parts  per  1000. 

In  a  third  article,  of  about  the  same  date  as  the  above,  Peritz 
(1908-9b)  submits  lecithin  estimations  on  the  serum  of  a  large 
number  of  cases  of  lues,  tabes,  paralysis,  and  other  nervous  affec- 
tions. The  figures  for  the  same  disease  vary  remarkably.  We  dis- 
cover nothing  characteristic  in  them. 

Peritz  (1910)  reported  further  work  of  a  similar  nature.  Both 
ingestion  and  injection  of  lecithin  in  syphilis,  tabes,  neurasthenia, 
and  the  normal  state  are  shown  to  increase  the  lecithin  in  the 
serum.  Lecithin  determinations  were  also  made  in  the  fat  of  the 
bone  marrow  of  cases  of  dementia  paralytica.  These  figures  varied 
from  0  to  4.21  percent  of  lecithin  in  the  fat. 


PHOSPHORUS  METABOLISM 


499 


«2 
<J 

W 

I— i 
M 

P  DO 

S  s 

W  <j 

■H  H 

^  »2 

A  HH 

£  O 

^  5    a 

ft  q  S 

OtJ     *h 

i-J  H 

w  fe  s 

woe 

^  OQ    « 

g  s  -a 

GO  y     N 

§  a  s 

M  H  S 

&  >*    « 

<=>  s  ^ 

S  w 

Q  O 

jz;  w 

^  (si 

H  «*! 

sz; 

i— i 

« 
o 
j 

m 
u 


"?  bo 

a;  -^ 
.9  a 

6  c4 

8^ 
ft 


S3 .2 


2.2 
M  o 


<jd 


fe  .a 

+J    7! 


2.2 
•8  m 
pq  o 


S3 .2 

-4->    W 

-sin 


2.2 

•2.2 
ffl  u 


COCO      "CO 


■»«coa5r-.ioiocoT*(a»oom<Noa  ■  •■— icornco 

COOO^CMCMOOOiCOt^CJiOO.— ICCO  •  .-31CMaO<-H 

lOiniowcoorfWrHHOTHW  •  .1 — tc^j  ^ . — i 

ooooooooooooo  •  -oooo 


•  IO  — 100-flicOTti 
•CM— i  CM  CO  COO 

•oooooo 


ooooooooooooo 


•oooo 


•oooooo 


.i-lt>  •  «-> 

•tf*  •  ■« 

•COCX)  ••— I 

•  oo  -o 

•oo  ■© 


i— iTjtcqsoeMosooor^t^eNisocn    •    .cocdocm 


•  r-c---»CMCiocD 


CMOOoioiocooocMr^oocMco    .    -cMCMr-oo    .    -r^r-coioor- 


COlOlOOT*r-1t^TJ1CMCOCMCMr 

ooooooooooooo 


•  oooo 


•  co  oo  in  -rr  io  m 
•oo  oooo 


ooooooooooooo 


•oooooo 


oo 
oo 


•  OSr-    .00 


io  c^  cm  co  it- ooco  cm  coior-oo  cm    .    .-ticM-tno 


— tt--COCMt— IO 


QO  CO  CM  00  C--  IO  03  TJ1  O t--  CD  CO  O      °     .COlOCOCft      •     •  CO  00  ""*  03  •*  CM 
r- <  ^H  CO  i— I  CO  CM  i— I  CD  Tfl  0O  CO  CO  •*      •      •lOTftlOCO      -      -  02  03  O0  t>  00  IO 


HrtrtOOOHOOOOOO 

oo'ooooooooooo 


•  oooooo 
•oooooo 


•oco    .-SI 

.TJIO      *CO 
•IT-CO      -IO 


O©   CUNNMOL^OOOOOOOOOOffJ 


tmco  oooooot^ocoro 


-1CDI--C0-3110CO~H 


-     ..     <i— IC0  03COOr--CDCOr— lOOOIOiT-CO 
©CM    jhOOOOO'*CMti(CMCOt-ICOCO'"*i<MO 

o'o£Ho'oo'oooooooo'ooo*orH 


•  oooooo 

•CMOCOCOCOO 

•  osoococoio-^ 


•OOOOOO 


2.2 
•B  m 
<u"E 


u  ai 


2.2 

a.  2 

<D   I* 

pq  u 


a>©  n  iccB'Jooootiioooo  ooo  moooooo 

U-rH    0  CM  CM  -*  IO  —4  CO  -31  O  IO  -*  Cn  IO    U-H-*    OOO^HOTCOCoS 

rt°.  ti-  2SSSSS2SSS22  SJ!2  «22£  -  =  = 

&_,0  ^,«  ©©©©©©©©©©O©  j_,00  i^,©©©©©© 

HoH  ooo'ooo'oooooofHoo^oooo'oo 


NO     -CO 

©-31       'Ttl 
CMCO     'CM 


.NOjtMBOcgruNooainooomooomooiNMTiit-. 

.— -li-H-31^t003'^1lOCOCO©-31t^©-31iOt^.'*+<OiCO©COCOCMi— I 
•COCOTtlCOCOCOCOCMCMCO— HCOCOCMCMCMCOCOCMCMCOCMCMCOCM 

•  ooo<oo<oo>ooooooooooo^oooooo 
•oooo'o'oo'ooo'ooo'o'o'ooo'o'oooooo 


h  o> 


<Jt3 


2.2 


.CMOT     -CM 


CMOOOO-*-*CDCC>COCOiO-*-*1CSCMCCroiNCOCOC^TtlC73C-J03C>a!IO 
-31CC0^HCMCOr^O5CO-*1COTf1'— last-COt—OO^lCOCOOi— HCD"^0010t— 1 
CMC-JNCN10ai>JCMCMCM0qCMCOCM>-WCMCMC>)CNCMCJOQC^CMC^CMC^ 

ooooooo'ooooo'o'ooooooooooooocJ 


g:  :  :  :   feg:   fe§: 


:  i  :  s  z  fes 


uS  ™ 
y -^  a 

C  u  (i>  oT 

*»  ft.9  -r 
-!»  «     b 

.2 .2  ft* 


S'E'E  fi 
■axs-a-d 

Pi  ty  Hi  53- 


<N: 

-* 
m 

P 

'3 

a; 

n 

h 

CJ 

CO 

- 

500  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Bornstein's  (1909)  studies  of  the  blood  in  cases  of  progressive 
paralysis  give  us  the  following  values  for  the  lecithin  content  of 
blood  serum.  Normal  serum  is  said  to  contain  2.0-2.4  parts  lecithin 
per  1000  parts  of  serum ;  and  serum  from  these  subjects  having  pro- 
gressive paralysis  contained  3.5,  2.7,  2.8,  2.8  and  2.7  parts  and  that 
from  one  subject  having  tabo-paralysis,  2.9  parts  per  1000. 

Glikin  (1909c)  determined  lecithin  in  the  blood  in  polycythaemia 
rubra  megalosplenica,  a  disease  in  which  the  blood  contains  an  abnor- 
mal amount  of  iron.  As  compared  with  Abderhalden's  figure  of 
2-3  gm.  lecithin  per  1000  c.c.  of  blood  (for  mammalia),  Glikin's 
determinations  of  5.226  and  5.041  gm.  per  1000  c.c.  in  this  disease 
make  it  appear  twice  as  high  as  the  normal. 

Takemura  (1910)  determined  total  phosphorus  in  the  blood 
serum  of  cases  of  syphilis  and  cancer,  and  from  subjects  in  the  nor- 
mal state.  The  averages  for  syphilis  and  cancer  are  both  a  little 
higher  than  for  the  normal  subjects,  though  there  is  nothing 
characteristic  in  the  individual  figures. 

Kimura  and  Stepp  (1911)  note  Peritz's  finding  of  a  high  leci- 
thin content  of  the  blood  serum  in  lues,  tabes  and  paralysis,  and  at 
the  same  time  an  elimination  of  lecithin  in  the  feces,  from  which  he 
concluded  that  these  diseases  involve  an  impoverishment  of  the 
body  in  lecithin.  Kimura  and  Stepp  continued  this  study,  and 
reported  ether-soluble  phosphorus  determinations  on  the  serum 
of  36  cases  of  various  diseases,  with  results  in  grams  of  lecithin,  per 
1000  c.c.  serum,  as  follows:  High  values,  pneumonia  2.10,  typhoid 
convalescent  2.0,  optic  neuritis  2.15,  diabetes  mellitus  2.36-2.60,  lues 
(spinal)  2.27,  tabo-paralysis  1.97;  low  values,  Basedow's  disease  0.9, 
chronic  nephritis  0.60-1.1,  and  cardiac  insufficiency  with  chronic 
nephritis  0.5. 

According  to  Sawadski  (1911),  the  normal  content  of  blood 
corpuscles  in  organic  phosphorus  is  0.8  per  1000,  that  of  the  serum 
0.17  per  1000,  that  of  the  inorganic  phosphorus  in  the  erythrocytes 
is  0.6-0.7  percent,  in  serum  0.12-0.14  percent.  In  diseases  which 
are  accompanied  by  an  increased  acid  content  of  the  blood  the 
amount  of  inorganic  phosphorus  increases.  In  infectious  diseases 
the  amount  of  organic  phosphorus  of  the  serum  is  less ;  in  uraemia 
both  kinds  of  phosphorus  rise.  The  partition  of  both  the  organic 
and  the  inorganic  phosphorus  between  serum  and  erythrocytes  is, 
under  normal  conditions,  1:5.  By  pathological  processes  only  the 
quotient  for  organic  phosphorus  changes. 

The  following  are  the  averages  of  Pusanow's  (1911)  analyses 
of  the  mineral  part  of  normal  blood  and  that  from  sufferers  from 
arteriosclerosis. 


PHOSPHORUS  METABOLISM 


501 


MINERAL  DETERMINATIONS  ON  BLOOD  IN  NORMAL  AND 
PATHOLOGICAL  CONDITIONS  (Pusanow,  1911)  Percent 


Condition 


Normal 

Arteriosclerosis. 


Number 
of  cases 


P2O5 


0.33 
0.43 


SO3 


0.67 
0.85 


0.80 
0.81 


CaO 


0.01 
0.17 


Burger  and  Beumer  (1913a)  found  cholesterol  and  lecithin 
values  in  blood  serum  higher  in  diabetic  lipaemia  and  cholaemia 
than  in  other  diseases  studied. 

LECITHIN  THEEAPT  IN  DISEASES   OF  THE  BLOOD 

Jolly  (1887a)  and  others  have  expressed  the  belief  that  pover- 
ty of  the  food  in  phosphorus  compounds  may  contribute,  as  well  as 
lack  of  iron,  to  the  cause  of  anaemia.  Whatever  the  facts  in  this 
matter,  we  have  a  number  of  reports  of  attempts  to  cure  anaemia 
by  phosphorus  compound  therapy. 

Kepinow  (1910)  found  that  the  lipoids  of  beef  blood  injected 
into  rabbits  which  had  been  rendered  anaemic  by  bleeding,  led  to  a 
greatly  accelerated  renewal  of  the  subject's  blood,  as  compared  with 
controls.     The  injection  of  lecithin  was  not  attended  by  like  results. 

Tonelli  (1898)  administered  lecithin  in  cases  of  anaemia  and 
chloranaemia,  and  noted  increase  in  body  weight  and  haemoglobin ; 
and  in  simple  anaemia,  increase  in  the  number  of  red  blood  corpus- 
cles. There  was  also  improvement  in  general  condition  and  in  the 
digestion. 

Muggia  (1898)  described  3  cases  of  anaemia  and  infantile 
athrepsia  which  were  treated  by  injection  of  lecithin  and  egg  yolk. 
The  blood  was  examined.  These  treatments  are  said  to  have  had 
nutritive  and  curative  value. 

F.  Levy  (1905)  reports  5  cases  of  anaemia  which  were  treated 
with  a  lecithin-containing  preparation,  "lecitogen."  This  medica- 
ment contained  a  variety  of  ordinary  nutrients,  and  0.94  percent  of 
lecithin.  A  general  increase  of  haemoglobin  and  erythrocytes  was 
noted  in  the  blood.  We  could  present  a  long  list  of  common  foods 
containing  very  much  more  lecithin  than  does  lecitogen.  The 
improvement  in  these  cases  was  probably  due  to  other  constituents 
of  the  preparation,  though  we  should  reserve  the  possibility  that 
lecithin  in  the  free  state  is  absorbed,  in  larger  part  as  such,  than 
is  the  phosphorus  of  combined  lecithin,  in  which  case  the  free  leci- 
thin might  be  more  useful  in  metabolism. 


502  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Lewin  (1905)  reports  results  of  blood  and  urine  examination 
with  8  patients  suffering  from  chlorosis  and  secondary  anaemia, 
under  treatment  with  bromlecithin.  The  5  cases  of  chlorosis  were 
said  to  be  pronounced  and  typical.  In  all  cases  there  was  evident 
improvement,  as  shown  by  increase  in  haemoglobin  and  in  the  num- 
ber of  erythrocytes,  as  well  as  in  favorable  changes  in  clinical 
appearance. 

Bergell  and  Braunstein  (1905)  administered  bromlecithin, 
containing  20  percent  bromine,  in  quantities  of  0.2  gram  3  times 
daily  to  anaemic  patients,  with  results  of  moderate  increase  in 
haemoglobin  and  red  blood  corpuscles.  Seven  patients  were  included 
in  this  study,  aggregating  13  periods  of  4  to  28  days.  The  drugs 
were  administered  in  pill  form. 

NUCLEIN  THERAPY  IN  ANAEMIA 

Jacob  and  Bergell  (1898)  studied  metabolism  in  various 
diseased  conditions  as  affected  by  a  nuclein  preparation  from  calf 
spleen.  This  extract  was  in  the  nature  of  a  "beef  tea."  It  con- 
tained in  1  c.c,  0.0083  gm.  N  and  0.0034  gm.  P205.  Of  the  phos- 
phorus 93  percent  was  organic,  35  percent  being  nuclein  phosphorus. 

Carefully  conducted  balance  experiments,  preceded  by  proper 
preliminary  periods,  showed  in  secondary  anaemia  an  increase  of 
leucocytes,  during  12  days,  from  3,680  to  11,250;  and  erythrocytes 
from  4,200,000  to  5,775,000  per  cubic  millimeter;  in  pernicious 
anaemia  there  was  no  improvement ;  in  anaemia  following  parturi- 
tion the  erythrocytes  increased  during  6  days  from  1,450,000  to 
2,625,500,  the  leucocytes  remaining  constant,  while  the  haemoglobin 
increased  from  20  percent  to  31  percent ;  in  anaemia  with  uterine 
cancer  the  erythrocytes  did  not  increase,  but  the  leucocytes 
increased  during  8  days  from  3,100  to  6,800 ;  in  chlorosis  there  was 
an  increase  during  9  days  from  325,000  to  4,100,000  erythrocytes, 
and  at  the  same  time,  a  decrease  from  13,900  to  5,100  leucocytes. 

In  an  experiment  with  a  woman  21  years  old,  under  carefully 
regulated  conditions,  the  authors  found  that  the  nuclein  preparation 
was  well  absorbed,  but  that  it  was  without  perceptible  influence  on 
metabolism. 

The  authors  consider  the  more  striking  results  of  their  investi- 
gation to  be  the  diuresis  caused  by  spleen  extract  in  secondary 
anaemia,  the  thorough  absorption  of  the  nuclein  phosphorus,  the 
improvement  in  blood  conditions,  and  the  improved  phosphorus 
retention.  They  recommend  the  administration  of  nuclein-contain- 
ing  diet  in  the  treatment  of  all  organisms  suffering  from  a  loss  of 
phosphorus. 


PHOSPHORUS  METABOLISM  503 

CALCAREOUS  DEGENERATION 

Almost  nothing  on  this  important  subject  has  come  to  our 
attention  in  which  the  participation  of  phosphorus  has  received 
mention. 

0.  Klotz  (1905)  has  studied  the  process  of  calcareous  degenera- 
tion, and  concludes  that  the  first  step  in  the  process  is  cloudy  swell- 
ing, or  coagulation  necrosis ;  following  this  there  are  fatty  changes 
in  the  cells;  and  soaps  of  sodium,  potassium,  and  presumably 
ammonium,  are  to  be  detected.  These  soaps  combine  with  calcium 
salts  in  solution  in  the  body  fluids.  Later,  judging  from  the  fact 
that  phosphate  and  carbonate  of  lime  are  formed,  and  the  deposits 
give  no  reaction  for  fats,  the  fatty  acid  moiety  of  the  calcium  soap 
is  replaced  by  the  more  powerful  phosphoric  and  carbonic  acids. 

Barille  (1910)  advances  the  theory  that  atheromatic  deposits 
of  calcareous  material  are  due  to  decreased  oxidation,  the  reduced 
amount  of  carbon  dioxide  being  insufficient  to  keep  the  phosphates 
in  solution,  in  which  connection  might  be  mentioned  the  work  of 
Flesch  (1876),  who  determined  that  all  of  the  constituents  of  bone 
are  dissolved  from  bone  powder  suspended  in  water  into  which 
carbonic  acid  was  led,  the  whole  being  kept  at  body  temperature. 

A  peculiar  pathologic  retention  of  phosphates  is  described  by 
Vannini  (1911)  under  the  name  of  "kalkgicht."  Calcium  salts 
were  deposited  in  quantity  beneath  the  skin.  The  balances  of 
sulphur  and  chlorine  were  negative ;  other  mineral  elements,  especi- 
ally the  alkaline  earths  and  phosphorus,  were  stored. 

CANCER 

The  concensus  of  opinion  has  it  that  in  cancer  the  phosphorus 
of  the  urine  is  often,  though  by  no  means  always,  increased,  usually 
coincidently  with  increased'  nitrogen  elimination,  both  resulting 
from  tissue  destruction.  Under  circumstances  which  result  in 
starvation  the  relative  amounts  of  the  urinary  constituents  show 
that  the  bones  as  well  as  the  soft  parts  are  contributing  to  the 
phosphorus  excretion.  The  metabolic  effects  of  cancer  are  the 
consequences  of  its  growth,  of  whatever  nature  these  effects  may  be 
as  determined  by  the  parts  involved,  and  of  the  disturbed  state  of 
nutrition,  especially  due  to  undernourishment.  There  is, 
therefore,  nothing  characteristic  in  general  phosphorus  metabolism 
in  cancer. 

Among  the  many  investigations  of  metabolism  in  cancer  the 
following  have  to  do  especially  with  phosphorus  compounds :  Miiller, 
F.,  1889 ;  von  Recklinghausen,  1891 ;  Apolant,  1893 ;  von  Moraczew* 
ski,  1895b,  1897c;  Braunstein,  1903-4;  and  Takemura,  1910. 


504 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


Hannes  (1906)  reports  results  from  the  subcutaneous  injection 
of  2-percent  sodium  nucleate  solution  in  51  cases  of  uterine  cancer, 
the  same  being  administered  in  50  c.c.  quantities  10-16  hours  before 
operation  for  extirpation  of  cancer.  The  author  was  of  the  opinion 
that  this  treatment  increased  resistance  to  infection. 

Goodman  (1912)  finds  a  nuclease  in  carcinomatous  tissue  and 
considers  its  possible  connection  with  the  malignancy  of  carcinoma. 

Volter  (1913)  found  phosphatid  and  protein  phosphorus  lower 
in  carcinomatous  liver  tissue  than  in  neighboring  normal  tissue, 
the  nitrogen  and  fatty  acid  contents  showing  no  differences. 

Robin  (1911,  1913a)  reports  that  cancerous  portions  of  the 
liver  contain  more  inorganic  matter,  including  phosphorus,  than 
healthy  portions,  the  slower  the  cancerous  growth  the  greater  the 
accumulation  of  inorganic  matter. 

Robin  (1913b)  found  that  variations  in  urinary  phosphorus  in 
cases  of  cancer  were  not  characteristic  of  the  disease,  but  were  vari- 
able in  accord  with  secondary  conditions,  especially  the  amount  of 
food  consumed,  the  extent  of  the  tissue  destruction,  and  the  charac- 
ter of  the  tissues  involved.  He  does  not  in  this  latest  work  find 
retention  of  phosphates  in  cancerous  tissue. 

CARIES 

Mosetig-Moorhof  (1885)  found  no  dissolving  of  bone  after  the 
application  of  lactic  acid  for  the  purpose  of  destroying  fungous 
granulation  in  a  case  of  caries  fungosa. 

Conrad  Cohn  (1889)  made  comparative  analyses  of  normal 
dentine  and  such  as  had  been  affected  by  caries.  The  results  make 
it  appear  that  the  changes  wrought  by  the  disease  are  such  as  to 
increase  the  percent  of  both  water  and  organic  matter,  while  the 
percent  of  phosphates  and  carbonates  decreases.  The  calcium  and 
magnesium  phosphates  each  lose  fully  two-thirds,  and  the  calcium 
carbonates  about  four-tenths.  The  table  gives  the  mean  results  of 
the  analyses. 

COMPARISON  OF  THE  DENTINE  OF  NORMAL  AND  DISEASED  TEETH 

Cohn  (1889)— Percent 


Teeth 

Water 

Organic 
matter 

Lime 
phosphates 

Lime 
carbonates 

Magrnesia 
phosphates 

4.27 
10.91 

28.39 
66.38 

52.902 
14.470 

12.93 

7.92 

1.08 

0.35 

These  figures  show  a  phenomenal  increase  of  organic  material 
and  decrease  of  salts,  especially  calcium  phosphate. 


PHOSPHORUS  METABOLISM 


505 


Gassmann  (1908)  noted  that  the  lime  content  of  wisdom  teeth 
is  high,  and  of  the  teeth  of  the  dog  lower  than  in  man,  which  may  be 
a  reason  that  man's  teeth,  and  especially  the  wisdom  teeth,  yield 
to  caries  more  easily  than  the  dog's.  There  is  indication  also  that 
a  high  content  of  organic  matter  increases  the  general  resistance. 
The  analyses  compared  are  as  follows : 

COMPARISON  OF  TEETH  WHICH  DECAY  EASILY  WITH  THOSE  WHICH 
DO  NOT  (Gassmann,  1908)— Percent,  Fresh  Basis 


Human  teeth 

Teeth  of 

Canines 

Milk  teeth 

60-year  teeth 

Wisdom  teeth 

dog 

Ca 

22.20 
29.78 
0.87 
40.98 
4.18 
0.41 
0.34 
0.61 

22.84 
29.59 
0.78 
40.64 
4.12 
0.37 
0.35 
0.54 

21.42 
30.25 
0.82 
41.10 
4.32 
0.24 
0.27 
0.61 

Mean  of  2 
computed 

18.51 
31.77 

0.80 
41.48 

5.17 

0.40 

0.33 

0.67 

Mean  of  2 
computed 

26.11 

27.42 

Mg. 

0.76 

PO4. 

39.07 

CO3 

4.50 

CI 

0.18 

K 

0.15 

Na 

1.00 

99.37 

99.23 

99.03 

99.13 

99.19 

8.09 

8.76 

8.27 

6.83 

11.03 

Gassmann  (1910)  reports  an  increased  magnesium  content  of 
the  teeth  in  caries,  and  expresses  the  idea  that  increase  of  magne- 
sium in  bones  bears  an  essential  relation  to  pathological  conditions. 

EFFECTS  ON  PHOSPHORUS  METABOLISM  OF  CASTRATION  AND 

OVARIOTOMY 

Pinzani  (1898)  studied  the  effects  of  spaying  on  the  metabolism 
of  a  dog.  The  food  was  the  same  before  and  after  the  operation. 
Total  nitrogen,  urea  and  phosphoric  acid  in  the  urine  were 
decreased,  as  also  was  feces  nitrogen. 

Schulz  and  Falk  (1899)  found  in  metabolism  experiments  on 
dogs,  before  and  after  spaying,  that  this  operation  did  not  lead  to 
increased  phosphorus  excretion. 

Mosse  and  Oulie  (1899)  studied  the  effects  of  spaying  on  phos- 
phorus elimination  with  dogs.  The  removal  of  both  ovaries  caused 
a  moderate  increase  in  urinary  phosphorus.  The  ingestion  of 
ovaries  caused  a  reduction  in  the  phosphorus  outgo.  Spaying  was 
without  result  on  the  excretion  of  total  nitrogen. 

S.  Neumann  and  Vas  (1902)  studied  metabolism  in  dogs  as 
affected  by  spaying  and  by  ovarian  preparations  administered 
before  and  after  spaying.  Complete  balance  data  were  obtained. 
Subcutaneous  injection  of  ovarian  preparations,  with  a  normal  dog, 


506 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


caused  increased  excretion  of  nitrogen,  calcium  and  phosphorus. 
After  spaying  there  was  increased  nitrogen  but  decreased  calcium 
and  phosphorus  outgo.  The  feeding  of  ovarian  preparations  to 
spayed  dogs  caused  increased  excretion  of  calcium  and  phosphorus, 
but  no  important  change  in  nitrogen  metabolism. 

From  the  following  determinations  on  the  tissue  and  bones 
Heymann  (1904a,  1904b)  concludes  that:  "I.  It  is  certain  that 
castration  of  healthy,  female  animals  results  in  no  continued  phos- 
phorus retention.  2.  A  decrease  in  the  phosphorus  content  of 
the  organism  follows  castration.  3.  This  decrease  seems  to  affect 
the  phosphate  of  the  body  tissue  as  well  as  that  of  the  skeleton.  4. 
The  lecithin  content  is  not  affected  by  castration." 

PHOSPHORUS  COMPOUNDS  OF  RATS  AS  AFFECTED  BY  OVARIOTOMY 

Heymann,  (1904a,  1904b) 


Tissues,  percents  of  dry 
substance 

Bones,  percent 
total  P2O5 

Bones  and 

tissues, 
total  P2O5 

- 

Lecithin 
P2O5 

Nuclein 
P2O5 

Phos- 
phate 
P2O5 

Total 
P2O5 

Fresh 
sub- 
stance 

Dry 

sub- 
stance 

Percent  of  total 
body  weight 

0.4760 

0.3242 
0.3608 

0.3229 
0.2881 
0.2638 
0.3404 

0.0559 

0.0649 
0.0979 

0.0517 
0.1027 
0.0536 
0.0540 

2.4479 

1.6490 
1.5430 

1.3671 
1.2147 
0.6616 

2.9798 

1.9830* 
2.0018 

1.7417 
1.6055 
0.9790 

21.2690 
18.1665 
17.0315 
17.5724 

19.3633 
18.9100 
11.5705 
14.9766 

24.0556 
22.8105 
19.2083 
19.9277 

22.3595 
22.2174 
13.1568 
16.7290 

1.9819 

1.2980 

" 

? 

" 

1.3795 

41  d.  after  castration 
51  d.     " 
88  d.     " 
126  d.     " 

1.4928 
1.3875 
0.8481 

? 

*  Apparently  erroneous  since  the  sum  of  the  figures  for  nuclein,  lecithin  and  phosphate 
phosphorus   is   2.0381  percent. 

McCrudden  (1907,  1910a)  castrated  two  dogs,  and  spayed  two, 
in  a  study  of  the  effects  of  these  operations  on  mineral  metabolism. 
Of  his  results  McCrudden  says  in  part,  "It  will  be  seen  that  they 
do  not  confirm  the  view  generally  held  that  castration  is  followed 
by  a  general  retention  of  material,  especially  the  mineral  elements 
of  the  body.  In  fact,  my  results  show  a  general  tendency  in  the 
other  direction." 

This  work  was  controlled  with  care.  The  doubtful  trend  of 
the  results  of  these  experiments  and  of  those  which  precede  it 
shows  that  the  facts  as  to  the  influence  of  spaying  and  castration 
on  phosphorus  metabolism  can  not  be  markedly  characteristic. 

See  also  Daniel-Brunet  and  Rolland  (1911a). 

COMPOSITION  OF  THE  HUMAN  BODY  IN  DISEASE 

A  considerable  number  of  analyses  of  organs  of  human  beings 
dead  from  disease  may  be  found  in  print,  but  little  is  usually  made  of 
them,  and  indeed  little  can  be  made  of  them,  for  the  analyses  of  the 


PHOSPHOEUS  METABOLISM 


507 


parts  can  hardly  be  expected  to  afford  much  of  a  measure  of  their 
functional  activity.  This  most  important  purpose  can  be  served  only 
by  a  consideration  of  the  products  of  the  activity  of  the  organs.  On 
this  account  we  have  transcribed  but  few  such  analyses.  Von 
Moraczewski  (1897a),  however,  has  published  a  very  nice  series  of 
analyses  of  the  organs  of  human  beings  who  had  died  from  various 
diseases,  and  we  copy  these  and  his  discussion,  practically  complete, 
as  an  example  of  the  sort  of  evidence  which  may  be  obtained  from 
such  studies.  This  method  of  investigation  of  pathology  is  not 
entirely  without  value,  but  promises  comparatively  little. 

MINERAL  CONSTITUENTS  IN  HUMAN  ORGANS  AFTER  DEATH  FROM 
VARIOUS  DISEASES— Percent 


Nitro- 

Chlor- 

Phos- 

Phos- 

Cal- 

Dry 

in  titter 

gen 

ine 

phorus 

phorus 

cium 

Organ 

Disease 

in  fresh 

in  fresh 

in  fresh 

in  dry 

in  fresh 

Sex 

sub- 

sub- 

sub- 

sub- 

sub- 

stance 

stance 

stance 

stance 

stance 

Brain 

Pneumonia 

30.2 

1.159 

0.222 

0.291 

0.97 

0.007 

M 

* ' 

Carcinoma,  starvation 

14.5 

1.356 

0.153 

0.256 

1.76 

0.024 

F 

' ' 

Carcinoma,  anaemia 

7.15 

1.309 

0.213 

0.280 

3.98 

0.090 

F 

* ' 

Pernicious  anaemia 

18.4 

1.942 

0.152 

0.246 

1.33 

0.012 

F 

' ' 

Death  by  bleeding- 

21.3 

1.944 

0.145 

0.266 

1.25 

0.004 

M 

'  * 

Normal  state 

0.070 

0.002 

Heart 

Pneumonia 

16.9 

2.668 

0.142 

0.183 

1.08 

0.004 

F 

* ' 

Pneumonia 

14.0 

0.319 

0.149 

1.02 

0.009 

M 

" 

Carcinoma,  starvation 

12.0 

2.703 

0.157 

0.188 

1.57 

0.067 

F 

1 ' 

Carcinoma,  starvation 

13.7 

1.821 

0.153 

0.149 

1.09 

0.017 

M 

* ' 

Carcinoma,  anaemia 

8.8 

2.292 

0.170 

0.124 

1.38 

0.009 

F 

4 ' 

Carcinoma,  anaemia 

16.2 

2.237 

0.142 

0.093 

0.58 

0.001 

M 

*  * 

Pernicious  anaemia 

18.1 

2.758 

0.159 

0.160 

0.89 

0.005 

F 

* ' 

Pernicious  anaemia 

2L.9 

2.295 

0.189 

0.150 

0.68 

0.009 

M 

* ' 

Death  by  bleeding 

13.4 

2.319 

0.141 

0.121 

0.90 

0.002 

M 

' ' 

Normal  state 

36.0 

0.070 

0.203 

0.74 

0.007 

Spleen 

Pneumnoia 

16.6 

0.178 

0.237 

1.42 

0.002 

F 

' ' 

Pneumonia 

17.8 

4.237 

0.335 

0.331 

1.30 

0.001 

M 

" 

Carcinoma,  starvation 

17.9 

2.860 

0.215 

0.245 

1.36 

0.015 

F 

" 

Carcinoma,  starvation 

14.4 

2.646 

0.176 

0.188 

1.36 

0.003 

M 

l  e 

Carcinoma,  anaemia 

13.4 

2.464 

0.174 

0.218 

1.61 

0.004 

F 

1 ' 

Carcinoma,  anaemia 

19.2 

2.740 

0.198 

0.259 

1/81 

M 

*  ' 

Pernicious  anaemia 

11.8 

2.801 

0.259 

0.195 

1.65 

0.010 

F 

1  ' 

Pernicious  anaemia 

19.5 

0.186 

0.214 

1.09 

0.001 

M 

'  ' 

Death  by  bleeding 

8.3 

2.350 

0.140 

0.266 

2.14 

0.004 

M 

" 

Normal  state 

0.011 

0.132 

0.077 

Liver 

Pneumonia 

34.1 

4.071 

0.092 

0.189 

0.55 

0.001 

F 

' ' 

Pneumonia 

11.7 

1.584 

0.271 

0.932 

7.99 

0.004 

M 

" 

Carcinoma,  starvation 

17.2 

2.701 

0.191 

0.216 

1.25 

0.017 

F 

' ' 

Carcinoma,  starvation 

28.6 

2.022 

0.184 

0.180 

0.62 

0.001 

M- 

" 

Carcinoma,  anaemia 

38.6 

2.712 

0.174 

0.199 

0.51 

0.004 

F 

' ' 

Carcinoma,  anaemia 

17.4 

2.561 

0.153 

0.237 

1.35 

0.001 

M 

' ' 

Pernicious  anaemia 

9.1 

3.285 

0.216 

0.154 

1.69 

0.003 

F 

1 ' 

Pernicious  anaemia 

17.2 

2.565 

0.125 

0.217 

1.25 

0.017 

M 

' ' 

Death  by  bleeding 

4.3 

2.945 

0.209 

0.099 

2.30 

0.012 

M 

' ' 

Normal  state 

0.027 

0.388 

0.028 

Kidney 

Pneumonia 

9.4 

2.293 

0.188 

0.181 

1.92 

0.003 

F 

'  * 

Pneumonia 

12.2 

2.155 

0.392 

0.156 

1.28 

0.004 

M 

* ' 

Carcinoma,  starvation 

12.8 

2.291 

0.167 

0.131 

1.04 

0.012 

M 

' ' 

Carcinoma,  anaemia 

3.5 

1.913 

0.218 

0.178 

5.05 

0.003 

F 

" 

Carcinoma,  anaemia 

16.7 

1.793 

0.038 

0.214 

1.22 

0.001 

M 

'  * 

Pernicious  anaemia 

10.4 

2.404 

0.263 

0.153 

1.47 

0.008 

F 

' ' 

Pernicious  anaemia 

11.8 

1.933 

0.255 

0.166 

1.35 

0.009 

M 

'  * 

Death  by  bleeding- 

9.8 

2.421 

0.136 

0.161 

1.64 

0.003 

M 

Lungs 

Pneumonia 

12.4 

2.734 

0.191 

0.188 

1.51 

0.004 

F 

*  * 

Pneumonia 

9.9 

1.581 

0.246 

0.138 

1.39 

0.005 

M 

Blood 

Pneumonia 

0.276 

0.035 

0.003 

F 

• ' 

Pneumonia 

0.242 

0.032 

0.002 

M 

* ' 

Anaemia 

0.301 

0.008 

0.005 

F 

Anaemia 

6.96 

0.995 

0.300 

0.015 

0.21 

0.005 

M 

508  OHIO  EXPERIMENT  STATION:  TECHNICAL  BTJL.  5 

Discussion  by  author.  "While  the  phosphorus  in  the  main 
follows  the  variations  of  the  nitrogen,  it  is  the  opposite  with  the 
chlorine  and  the  calcium.  The  minimum  phosphorus  content 
accompanies  the  maximum  chlorine  content,  while  it  usually  corre- 
sponds with  the  minimum  of  nitrogen. 

"This  is  true  of  the  relations  under  different  causes  of  death  as 
well  as  for  the  individual  organs.  Of  the  organs  the  brain  only  is 
an  exception,  in  that  here  the  chlorine  parallels  the  phosphorus,  the 
nitrogen,  on  the  other  hand  increases  and  decreases.  In  the  other 
organs  as  well  as  in  the  different  bodies  there  is  the  antagonistic 
relation  of  chlorine  and  phosphorus.  I  must  emphasize  that  this 
is  true  only  for  most  cases  and  it  is  not  the  rule  throughout,  for  both 
parallelisms  and  irregularities  of  variations  in  the  series  of  figures 
are  observed. 

"The  relation  of  chlorine  to  phosphorus  seems  to  me  most  plain 
in  the  direction  mentioned.  The  parallelism  between  phosphorus 
and  nitrogen  is  less  plain,  for  one  observes  not  uncommonly  that 
the  amount  of  chlorine  runs  parallel  with  the  amount  of  nitrogen 
rather  than  the  amount  of  phosphorus.  In  general  in  the  case  of 
individual  organs  the  phosphorus  content  corresponds  rather  to  the 
nitrogen  content,  on  the  other  hand  in  the  individual  bodies  the 
phosphorus  and  chlorine  are  proportional  to  the  nitrogen,  which  can 
be  understood,  for  there  the  water  content  of  the  organs  plays  an 
important  part.  For  the  consideration  of  the  relations  of  the 
mineral  parts  with  one  another,  therefore,  the  series  of  numbers 
should  be  used  in  which  they  are  arranged  according  to  the  organs. 

"The  calcium  is  usually  present  in  so  small  an  amount  that  one 
cannot  form  a  correct  conception  as  to  its  relation.  It  gives  the 
impression  that  it  goes  parallel  with  the  chlorine  rather  than  the 
phosphorus.      Here  also  there  is  no  rule  shown. 

"There  is  therefore  no  clear  relation  of  the  mineral  parts  to  one 
another  and  to  the  nitrogen,  nor  is  there  any  more  a  difference 
between  the  different  bodies  established.  I  expected  to  find  a  typical 
relation  between  the  chloride  and  the  phosphorus,  as  it  is  so  often 
found  in  the  blood  in  life.  I  thought  the  anaemic  body  would  appear 
very  different  from  the  non-anaemic. 

"This  is  not  so.  There  is,  it  is  true,  a  chlorine  richness  of  the 
anaemic  body  as  compared  with  the  non-anaemic,  and  I  will  not 
neglect  to  emphasize  this ;  but  this  difference  in  chlorine  content  is 
far  less  marked  than  was  that  in  the  blood  in  life. 

"Still,  one  thing  is  made  fairly  certain  by  these  analyses:  the 
accumulation  of  chlorine  in  the  organs  and  the  impoverishment  of 
them  in  phosphorus  and  calcium.      All  known  analyses  of  organs 


PHOSPHORUS  METABOLISM  509 

up  to  this  time,  with  whatever  errors  may  have  been  made,  agree  in 
showing  a  low  chlorine  content — about  0.07%  computed  to  the  fresh 
substance.  Our  analyses  show  an  average  of  0.20%,  or  about  three 
times  as  much.  The  phosphorus  content  is,  according  to  the  differ- 
ent authors,  0.3-0.2%  ;  we  found  usually  less  than  0.2%.  In  normal 
organs  the  amount  of  phosphorus  is  3-7  times  as  great  as  the 
amount  of  chlorine;  with  our  cases  it  is  often  smaller  than  the 
amount  of  chlorine  and  but  seldom  larger,  usually  equal.  It  is 
especially  surprising  that  in  the  spleen  the  phosphorus  content 
seems  to  be  increased  and  in  the  liver  and  muscle,  decreased.  This 
signifies  that  the  organs  lose  chiefly  calcium  phosphate  and  retain 
only  nuclein  phosphorus.  The  storage  of  chlorine  is  caused  by  the 
organism  becoming  watery.  That  this  is  not  a  general  condition  of 
cadavers  is  shown  by  the  fact  that  the  figure  for  dry  substance 
usually  given  never  falls  below  30%.  Our  analyses  give  repeatedly 
figures  that  are  lower  than  20%.  The  large  number  of  determina- 
tions support  this  number ;  moreover,  it  has  long  been  known  that 
the  blood  may  in  anaemia  contain  only  8%  dry  substance  in  place  of 
the  usual  20%,  so  that  a  growing  watery  condition  of  the  organs  is 
nothing  unusual.  This  high  water  content  carries  with  it  the  high 
salt  content.  Further,  it  carries  with  it  that  the  water-soluble  salts 
are  principally  represented,  therefore  chlorides  and  soluble  phos- 
phates, not  calcium  phosphate.  C.  Schmidt  explains  the  high 
chlorine  content  of  the  blood  by  the  abundance  of  water  in  the  blood 
and  this  theory  has  in  recent  times  repeatedly  been  expressed,  to 
explain  the  retention  of  chlorine  in  anaemia  and  pneumonia. 

"There  seems  to  be  no  doubt  that  the  organs  become  watery 
in  life  in  certain  diseases.  Post  mortem  analysis  has  shown  this 
and  the  high  salt  content  may  well  be  explained  in  this  way. 

"In  consideration  of  my  former  experiments  I  think  I  am 
justified  in  a  further  conclusion.  I  think  that  the  organs  in  life 
change  under  all  kinds  of  disease  in  the  sense  that  they  become 
richer  in  water  and  in  salts,  but  that  the  blood  follows  the  relation- 
ship in  the  organs  only  when  it  itself  becomes  diseased— therefore 
only  in  anaemia.  This  is  the  only  way  to  explain  why  the  blood 
is  poor  in  chlorine  in  pneumonia  though  the  organism  retains 
chlorine,  and  why  with  like  chlorine  retention  by  the  organism  the 
blood  is  increased  in  chlorine  content  in  cases  of  anaemia. 

"The  taking  up  of  water  is  therefore  always  to  be  looked  upon 
as  a  sign  of  "disease  in  an  organ,  and  with  the  modern  theory  of  solu- 
tions many  of  the  remarkable  reactions  of  the  organism  may  be 
traced  to  this  'dilation'." 


510  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Dennstedt  and  Rumpf  (1905)  also  made  an  extensive  series  of 
analyses  of  human  organs  after  death  from  many  diseases. 

For  further  observations  see  discussions  of  the  specific  states 
or  diseases  of  interest. 

PHOSPHORUS  METABOLISM  IN  DIABETES   MELLITUS 

Diabetes  is  an  anomaly  of  metabolism  which  results  in  chronic 
excess  of  sugar,  especially  dextrose,  in  the  urine.  This  is  due  to 
excess  of  sugar  in  the  blood,  resulting  from  a  failure  of  the  liver  to 
convert  it  into  glycogen ;  as  caused  either  by  affection  of  the  glyco- 
genic center  in  the  brain,  or,  more  commonly,  by  the  absence  of  the 
internal  secretion  of  the  pancreas. 

In  advanced  cases  of  diabetes  there  is  evident  a  reduction  of 
oxidative  capacity  of  the  body  generally,  as  shown  by  the  appear- 
ance of  imperfectly  oxidized  metabolic  products,  largely  of  an  acid 
character,  in  the  urine.  The  acid  character  of  these  products  is  at 
least  largely,  though  probably  not  wholly,  the  cause  of  the  coma  in 
this  disease. 

In  severe  cases  there  is  further  disturbance  of  general  metabo- 
lism through  failure  of  digestion,  insufficient  intake  of  food,  and 
excessive  destruction  of  body  proteins. 

VonNoorden  found  an  excretion  of  calcium,  magnesium,  and 
phosphorus  much  in  excess  of  the  quantity  in  the  food,  and  of  phos- 
phorus much  in  excess  of  that  which  would  accompany  the  excreted 
nitrogen  in  the  soft  tissues;  and  concluded  that  this  was  due  to 
katabolism  of  bone.  A.  R.  Mandel  and  Lusk  (1904)  also  found 
phosphorus  elimination  excessive  in  diabetes.  VonNoorden  cites 
the  work  of  Gaethgens,  showing  that  when  there  is  no  acid-intoxi- 
cation the  excretion  of  calcium  is  normal,  and  the  work  of  Van- 
Ackeren  showing  that  the  bones  atrophy  as  a  whole,  though 
vonNoorden  is  of  the  opinion  that  these  phenomena  are  not  due 
simply  to  the  action  of  an  acid.  VonNoorden  also  has  shown  an 
increased  excretion  of  purin  bodies  in  such  severe  diabetes  as  results 
in  much  destruction  of  tissues.  He  cites  the  work  of  Gaethgens  and 
Kulz  showing  that  the  natural  parallelism  between  nitrogen  and 
phosphorus  of  food  and  excreta  usually  exists,  except  where  there  is 
acidosis.  Under  this  latter  condition  the  phosphorus  excretion,  as 
previously  noted,  becomes  supernormal  in  comparison  with  the 
nitrogen  outgo. 

Gerhardt  and  Schlesinger  (1899),  and  also  vonNoorden,  showed 
that  under  these  conditions  of  excessive  phosphorus  outgo  in  diabet- 
ic acidosis  the  ingestion  of  sodium  bicarbonate  served  greatly  to 
reduce  this  excess  excretion  of  phosphorus  in  proportion  to  nitrogen. 


PHOSPHORUS  METABOLISM  511 

Von  Moraczewski  (1897b)  found  in  one  case  of  diabetes  melli- 
tus, on  a  mixed  diet,  a  retention  of  nitrogen  and  chlorine  coincident 
with  a  loss  of  calcium  and  phosphorus,  the  phosphorus  loss  being 
nearly  three  times  as  great  as  the  calcium  loss.  The  phosphorus 
loss  was  32  percent  of  the  intake,  and  the  calcium  loss  11  percent  of 
the  intake.  On  an  animal  diet  containing  much  less  chlorine  and 
lime,  and  somewhat  less  phosphorus,  the  nitrogen  balance  remained 
positive,  but  the  chlorine  became  negative,  and  the  losses  of  phos- 
phorus and  calcium  were  increased.  Von  Moraczewski  thought 
that  the  lime  excretion  was  a  specific  symptom,  and  that  decreasing 
the  lime  in  the  food  decreased  the  sugar  excretion. 

A  year  later  von  Moraczewski  (1898a)  published  further 
balance  data  on  diabetes  mellitus.  When  added  to  a  mixed  diet, 
calcium  phosphate  (10  gm.  per  day)  seemed  to  cause  a  retention  of 
calcium,  perhaps  a  slight  increase  in  nitrogen  storage,  a  reduced  loss 
of  phosphorus,  and  a  reduced  excretion  of  sugar,  while  sodium 
chloride  (10  gm.  per  day)  appeared  to  have  an  unfavorable  influence 
on  nitrogen,  phosphorus  and  calcium  balances.  In  a  later  paper 
(1903-04)  von  Moraczewski  published  urine  analyses  from  three 
cases  of  diabetes  mellitus  on  various  diets.  The  ingestion  of  trical- 
cic  phosphate  was  said  again  to  have  reduced  the  sugar  excretion. 

That  the  increased  phosphorus  elimination  in  diabetes  is  not  a 
necessary  concomitant  of  the  pathological  elimination  of  sugar  is 
shown  by  the  work  of  Lepine  and  Maltet  (1902),  who  found  in 
phloridzin  diabetes  that  an  excretion  of  53  gm.  sugar  per  liter 
of  urine  did  not  increase  either  the  proportion  of  phosphorus  to  urea 
or  the  proportion  of  phosphorus  to  total  inorganic  salts  in  the  urine. 

Erben  (1907)  found  a  decreased  lecithin  content  of  the  blood 
in  diabetes  mellitus,  and  large  amounts  of  alkalis  and  calcium.  In 
a  later  study  he  found  the  blood  plasma  with  a  normal  lecithin 
content,  but  the  erythrocytes  with  lecithin  content  reduced. 

As  bearing  on  the  connection  of  the  pancreas  with  diabetes  an 
experiment  by  Falta  and  Whitney  (1908)  on  the  effects  of  extirpa- 
tion Of  the  pancreas  on  metabolism  in  the  dog  is  of  interest.  See 
table,  p.  512. 

The  extirpation  of  the  pancreas  greatly  increased  the  outgo 
of  all  the  constituents  studied,  the  increase  affecting  the  minerals 
more  than  the  nitrogen.  The  endogenous  uric  acid  elimination  was 
also  increased.  The  authors  did  not  consider  that  any  particular 
tissues,  especially,  took  part  in  the  katabolism,  but  that  there  was 
simply  a  much  exaggerated  hunger  metabolism. 


512 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


EXCRETION  OF   NITROGEN,   CALCIUM  AND   PHOSPHORUS  IN  THE 

URINE  OF  A  FASTING  DOG  BEFORE  AND  AFTER  EXTIRPATION 

OF  THE  PANCREAS 


Condition 

Dates 

Length 
of  period 

Hours 

Urine 

N 

Grams 

P2O5 
of  urine 

Grams 

CaO 
of  urine 

Grams 

June  5 

"    6 
..    ? 

"   8 

July  18-19 
"    19-20 
"    20-21 
"    21-22 

24 
24 
24 
24 

14 
24 
24 
24 

5.353 
3.394 
3.864 
4.208 

6.700 
11.834 
11.981 
12.012 

0.664 
0.588 
0.639 
0.726 

1.644 
1.275 
1.672 
2.100 

0.0175 

<  i           ' » 

0.0198 

"            " 

0.0246 

u            «t 

0.0366 

Pancreas  removed;  fasting 

0.0124 
0.1522 
0.0792 
0.0885 

PHOSPHORUS  METABOLISM  IN  ENDOARTERIITIS 

Hirschler  and  Terray  (1905)  studied  metabolism  as  affected  by 
endoarteriitis  in  a  patient  53  years  old.  The  data  show  that  bone 
dust  is  assimilable  by  the  human  being.  In  this  case  it  appears 
that  the  bone  dust  increased  the  storage  of  nitrogen,  calcium,  phos- 
phorus and  magnesium.  In  this  case,  as  also  in  these  authors' 
experiment  in  feeding  bone  dust  to  a  dog,  the  ingestion  of  this 
substance  reduced  the  urinary  phosphorus  and  increased  the  feces 
phosphorus.  In  this  case  of  endoarteriitis  the  patient,  weighing 
67.6-68.8  kg.,  was  not  maintained  in  phosphorus  equilibrium  on 
7.638  gm.  P205  daily  in  milk,  eggs  and  rolls ;  but  the  daily  addition 
of  3  gm.  bone  dust,  containing  38.8  percent  P205,  caused  an  increase 
of  the  total  phosphorus  of  the  ration  to  8.818  gm.  P205  daily,  and 
resulted  in  storage  of  this  element.  The  balance  data  are  as 
follows : 

AVERAGE  DAILY  METABOLISM  IN  ENDOARTERIITIS— Grams 


Period 
and 

days 

Food 
P2O5 

Urine 
P2O5 

Feces 
P2O5 

Balances 

Blood 

P2O5 

Percent 

P2O5 

N 

CaO 

MgO 

Diet 

I 
3  days 

II 

3  days 

III 

3  days 

7.638 
8.818 

2.763 
5.536 
4.580 

1.206 
3.224 
3.763 

-1.2071 

+0.475 

—1.491 
+1.61 

+1.3051 
+1.790 

-0.3861 
+0.240 

0.0379 

0.060 

0.066 

Mixed,    normal,  much 
meat. 

Mostly  milk,  also  eggs 
and  rolls. 

Same  as  above  plus  3 
gm.  bone  dust. 

(1)       The  balance  figures  take  into  account  matter  vomited  and  blood  drawn. 

FATTY  DEGENERATION 

As  a  result  of  a  very  great  number  of  investigations,  the 
current  belief  in  regard  to  the  nature  and  cause  of  fatty  degenera- 
tion, so  called,  is  that  this  condition  results  from  one  or  the  other  of 


PHOSPHORUS  METABOLISM  513 

two  processes,  or  from  both  acting  at  once:  (1)  it  may  be  caused 
by  infiltration  of  fat  from  outside  the  tissue,  a  process  which  may 
be  either  entirely  normal,  or  due  to  injury  to  the  cells,  and  which 
may  be  either  without  demonstrable  effect  on  the  functional  efficien- 
cy of  the  tissue,  or  which  may  proceed  to  such  an  extent  as  to  cause 
interference  with  metabolism,  and  even  the  death  of  the  cell 
nucleus;  or  (2)  it  may  result  from  the  release  of  the  tissue  fats  by 
the  autolysis  of  the  proteins  with  which  they  are  chemically  com- 
bined, a  process  which  may  greatly  increase  the  visible  fat  without 
any  increase  in  the  amount  actually  present;  or  (3)  both  of  these 
factors  may  operate  simultaneously. 

That  the  lecithin  of  the  tissues  does  not  constitute  a  source 
of  the  fat  which  becomes  demonstrable  in  the  second  type  of  fatty 
degeneration  has  been  shown  by  Lusena  (1903)  and  Rubow  (1905), 
who  found  that  the  lecithin  of  cells  remains  practically  normal  even 
in  extreme  fatty  degeneration.      See  also  Krehl  (1893). 

Lepine  (1901)  found  that  some  fatty  human  livers  contained 
as  much  as  3  percent  of  lecithin,  on  the  fresh  basis  (15  percent  on 
the  dry  basis),  and  that  in  such  cases  the  urine  contained  much 
more  glycerophosphoric  acid  than  normal.  He  suggests  this  abund- 
ance of  incompletely  oxidized  phosphorus  of  the  urine  as  an  indica- 
tion of  a  fatty  condition  of  the  liver. 

Lepine  and  Eymonnet  made  a  study  of  the  glycerophosphoric 
acid  of  over  100  cases  of  different  diseases.  In  fatty  degeneration 
of  the  liver  in  tuberculosis  he  found  the  amount  of  glycerophos- 
phoric acid  in  the  urine  to  be  1.0-1.8  percent  of  the  amount  of  the 
nitrogen,  as  compared  with  0.15-0.30  in  the  normal  man.  See  also 
F.  Munk  (1908). 

Balthazard  (1901b)  found  in  a  human  liver  a  high  lecithin  con- 
tent associated  with  fatty  degeneration,  and  later  made  the  same 
observation  on  goose  livers.  The  amounts  of  phosphorus  in  the 
goose  livers  differed  rather  widely,  however,  a  fact  which  Balthazard 
thinks  is  probably  due  to  their  being  taken  at  different  stages  of  the 
fattening  process,  which  he  thinks  occasions  first  a  surcharging  of 
the  liver  with  lecithin,  and  second  a  replacement  of  the  lecithin  with 
fat,  this  latter  process  being  accompanied  by  an  excessive  elimina- 
tion of  glycerophosphoric  acid  in  the  urine  (Lepine). 

Balthazard  (1901a)  considered  that  the  lecithin  content  of  the 
liver  increased  in  a  number  of  pathological  states,  tuberculosis,  diph- 
theria, phosphorus  poisoning,  experimental  typhus  intoxication, 
inanition  and  uremia.  The  determinations  of  the  normal  lecithin 
content  of  the  liver  of  the  various  animals  involved  were  not  shown 
to  be  adequate  in  number. 


514  OHIO  EXPEKIMENT  STATION:  TECHNICAL  BUL.  5 

Balthazard  thinks  that  a  large  part  of  the  hepatic  lecithins 
comes  from  the  destruction  of  leucocytes  of  the  circulating  blood. 

Satta  and  Fasiani  (1910)  conducted  autolysis  experiments  with 
liver  which  are  not  without  interest  in  this  connection.  They 
found  that  the  addition  of  lipoids  increased  autolysis  of  the  liver, 
as  measured  by  the  amount  of  nitrogen  which  passed  into  solution 
in  a  given  time.  This  action  is  most  pronounced  if  the  liver  of  a 
starved  animal  is  used  for  the  autolysis. 

A  vast  extent  of  evidence  is  at  hand  on  the  fatty  degeneration 
of  phosphorus  poisoning,  many  hundreds  of  investigations  on  this 
one  subject,  but  its  value  in  assisting  us  to  an  understanding  of 
other  conditions  is  not  sufficient  to  warrant  us  in  undertaking  the 
great  labor  of  its  organization.  We  are  obliged,  therefore,  as  a 
practical  measure,  entirely  to  omit  it  from  this  consideration. 

PHOSPHORUS  METABOLISM  IN  FEVER 

In  fever  there  is  generally  reduced  alkalinity  of  the  blood  as  a 
result  of  the  formation  of  various  compounds  of  acid  character, 
especially  the  acetone  bodies, — acetone,  diacetic  acid,  and  /?-oxy- 
butyric  acid.  The  first  two  are  oxidation  products  of  the  last,  and 
in  the  disordered  oxidation  of  fever  they  naturally  appear  in  the 
order  mentioned,  and  disappear  in  the  reverse  order.  Their  form- 
ation is  thought  to  be  due  to  inanition  and  to  fat  destruction. 
Accompanying  these  acid  products  of  disordered  oxidation  is  an 
increase  in  the  urinary  ammonia.  In  many  fevers  retention  of 
chlorine  takes  place,  but  not  in  all,  by  any  means,  as  for  instance 
in  malaria.  In  convalescence  there  is  usually  a  negative  chlorine 
balance.  In  general,  the  elimination  of  chlorine  in  fever  seems  to 
be  reciprocal  to  the  elimination  of  phosphorus,  though  the  phospho- 
rus outgo  is  more  variable  than  the  chlorine  retention.  To  what 
extent  the  retention  of  chlorine  is  necessitated  by  the  phosphorus 
outgo,  in  order  to  maintain  the  isotonicity  of  the  blood,  has  not  been 
determined. 

Edlefsen  (1882)  observed  that  in  acute  stages  of  fever  the 
elimination  of  phosphorus  in  proportion  to  nitrogen  was  subnormal, 
and  explained  this  as  due  to  retention  for  the  growth  of  white  blood 
corpuscles. 

Rem-Picci  and  Bernasconi  (1894)  studied  phosphorus  excretion 
in  malarial  fever.      Some  of  their  observations  are  as  follows : 

The  total  phosphorus  excreted  in  24  hours  is  frequently  above 
normal.  With  increase  of  temperature  above  normal  there  occurred 
an  increase  in  the  urine,  and  at  the  same  time  a  decrease  of  urinary 
phosphorus,  even  to  5  mg.  P,O0  per  hour,  independently  of  the  diet. 


PHOSPHORUS  METABOLISM  515 

There  is  a  post-febrile  increase  of  urinary  phosphorus  compensating 
for  the  febrile  decrease.  Phosphaturia  frequently  follows  quinine 
treatment  of  the  fever.  There  was  established,  however,  no  definite 
influence  of  the  quinine  on  phosphorus  elimination.  These  results 
the  authors  attribute  to  an  inability  of  the  kidneys  during  fever  to 
excrete  the  phosphates  formed. 

Schwartz  (1895)  made  a  special  study  of  phosphorus  metabo- 
lism in  pneumonia,  in  balance  experiments  covering  4-7  days,  with 
three  subjects,  beginning  in  each  case  before  the  crisis  of  the 
attack,  and  continuing  until  2-3  days  after  the  critical  point. 
Schwartz  found  in  each  case  a  negative  phosphorus  balance.  The 
phosphorus  outgo  underwent  a  marked  decrease  beginning  before 
and  extending  through  the  crisis,  and  becoming  normal  soon  there- 
after. Schwartz  says,  "The  retention  of  phosphorus  belongs  in  no 
way  to  the  character  of  pneumonia;  still  less  to  fever  diseases  in 
general." 

Chlorine  was  retained  during  the  fever,  the  outgo  increasing 
suddenly  after  the  crisis.  Schwartz  observes  a  general  and  consis- 
tent reciprocal  relation  between  the  sodium  chloride  and  the  phos- 
phorus of  the  blood  serum  in  a  number  of  diseases;  that  is,  the 
higher  the  sodium  chloride,  the  lower  the  phosphorus.  He  believes 
that  this  is  caused  by  a  disturbance  of  osmotic  pressure  through 
decrease  of  phosphorus  in  the  tissues,  and  neither,  as  does  Terray, 
that  the  retention  of  choline  is  the  initiating  factor,  nor,  as  does 
Edlefsen,  that  the  increased  phosphorus  outgo,  after  the  crisis  may 
be  due  to  leucocytosis,  since  Limbeck  has  shown  that  this  is  no 
longer  present  after  the  crisis. 

Kuhnau  (1896-7)  studied  the  urine  in  a  case  of  malaria.  On  ten 
consecutive  days  the  phosphorus  (P,05)  excretion  in  the  urine  was 
as  follows,  in  grams,  2.131-0.201-1.746-0.231-1.695-0.437-1.876-1.594- 
2.13  and  1.84.  The  low  figures  were  from  days  of  fever,  when 
there  was,  at  the  same  time,  an  increased  excretion  of  uric  acid  and 
purin  nitrogen.  Kuhnau  raises  the  question  of  the  possibility  of 
this  being  due  to  the  use  of  phosphates  in  synthesis  by  the  parasitic 
organisms.  The  food  of  the  patient  contained  1.50-2.2  gm.  P205 
per  day. 

Von  Moraczewski  (1896)  published  many  urine  analyses  from 
which  he  concluded  that  in  acute  fever  diseases  there  is  little 
chlorine,  much  sulphur,  phosphorus  and  nitrogen,  and  an  especial 
increase  of  phosphate  earths,  as  in  anaemia.  In  diseases  where  there 
is  fever,  the  urine  shows  relatively  the  same  changes  as  the  blood, 
while  in  anaemia  opposite  relations  to  those  in  fever  exist. 


516  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Kalinin  (1897)  studied  urinary  phosphorus  excretion  in  three 
rabbits  and  a  dog  during  the  latent  period  of  fever  following  inocu- 
lation with  broth  cultures  of  B.  pyocyaneus  and  B.  diphtheriae.  The 
oxygen  consumed,  and  carbon  dioxide  excreted  were  decreased  in  the 
latent  period ;  both  increased  as  the  fever  rose.  In  the  latent  period 
the  urinary  nitrogen  and  phosphorus  were  low,  but  both  increased 
with  rise  in  temperature,  though  still  subnormal  during  the  first 
hours  of  the  fever. 

Ceconi  (1898)  also  noted  that  both  organic  and  inorganic 
phosphorus  in  the  urine  decrease  in  relation  to  nitrogen  in  high 
fever. 

Setti  (1898)  found  urinary  phosphorus  not  much  altered  in 
broncho-pneumonia. 

Hitzig  (1898-9)  studied  urinary  excretion  in  6  cases  of  malaria. 
The  hourly  excretion  in  grams  of  P,05  for  these  cases  on  the  fever 
days  and  the  fever-free  days  are  as  follows: 

Fever  day 0.0167 

Case  I  Fever-free  day    0.0475 

TT  Fever  day 0.0060 

Case  11  Fever-free  day    0.0598 

rs»<!f>  TTT  Fever  day  0.0016 

^a  Fever-free  day .0.0398 

Case  IV  Small  variations  for  fever-  and  fever-free-periods 

r        v  Fever  day  0.0067 

Case   v  Fever-free  day 0.0234 

Case  VI        Fever  day  0.0042 

Fever-free  day 0.0699 

The  relation  of  P205  to  nitrogen  in  the  urine  was  also  character- 
istic, as  follows : 

P205:N  P205:N 

Fever  After  Fever 

Case  1 1.60:100 6.4:100 

Case  2 0.46:100 8.3:100 

Case  3   0.78:100 14.9:100 

Case  4 0.58:100 5.7:100 

Case  5 2.45:100 5.5:100 

Case  6 0.36:100 8.8:100 

The  total  nitrogen  and  ammonia  increased  during  fever,  but 
the  proportion  of  ammonia  nitrogen  to  total  nitrogen  increased  five- 
fold. The  sodium,  potassium  and  chlorine  of  the  urine  greatly 
increased  in  fever,  the  phosphates  varying  in  the  other  direction, 
the  decrease  of  the  phosphates  beginning  before  the  onset  of  the 
fever. 

Paton,  Dunlop  and  Macadam  (1899)  studied  phosphorus  excre- 
tion in  the  urine  of  female  dogs  as  affected  by  diphtheria  infection 
(controlled  by  antitoxin),  and  by  fast.  As  an  average  of  results 
from  four  experiments  the  authors  found  a  normal  daily  urinary 


PHOSPHORUS  METABOLISM  517 

excretion  of  1.63  gin.  P205,  during  the  fast  1.35  gm.,  and  during 
fever  1.24  gm.  P205 ;  the  percent  of  P205  relative  to  total  nitrogen, 
21.9  normal,  25.7  in  fast,  and  17.9  in  fever.  These  figures  they 
consider  to  show  that  in  fever  nuclein  compounds  are  less  rapidly- 
decomposed  than  others  containing  a  lower  proportion  of  nitrogen  to 
phosphorus,  or  else  that  katabolic  phosphorus  is  retained. 

Von  MoraczewsM  (1899)  studied  urinary  excretion  in  various 
febrile  conditions.  Considering  the  course  of  an  attack  of  fever  as 
divisible  into  six  periods,  von  MoraczewsM  observed  the  interrela- 
tionship of  urinary  constituents,  and  plotted  curves  to  ishow  the 
initial  rise  in  chlorine  and  fall  in  phosphorus,  followed  by  a  more 
protracted  fall  in  chlorine  and  rise  in  phosphorus  elimination,  and 
then  an  increase  of  chlorine  and  decrease  of  phosphorus  to  the 
normal.  Variation  in  these  two  constituents  was  always  in  the 
opposite  direction. 

At  a  later  date  von  MoraczewsM  (1902)  published  observations 
on  a  case  of  acute  pneumonia  in  a  man  from  whom  the  spleen  had 
been  removed,  because  of  tumor,  seven  months  previously.  With 
rise  in  temperature  the  white  blood  corpuscles  fell  from  70000  to 
8000,  to  rise  again  when  the  temperature  began  to  fall,  and  finally 
to  return  to  the  normal  level.  With  rise  of  number  of  white  blood 
corpuscles  was  associated  a  parallel  decrease  in  urinary  phosphorus 
from  0.833  to  0.041  gm.  P205  daily,  and  associated  with  this,  a  cor- 
responding decrease  of  potassium  elimination.  Calcium,  however, 
varied  in  the  reverse  direction.  This  behavior  of  the  phosphorus 
excretion  is  attributed  to  the  increased  requirement  of  phosphoric 
acid  by  the  increased  formation  of  white  corpuscles;  while  the 
increased  elimination  of  calcium  is  perhaps  to  be  explained  by  an 
increase  in  the  katabolic  processes  in  the  bone  marrow  associated 
with  the  growing  number  of  white  corpuscles  in  the  blood. 

Von  MoraczewsM  (1900a)  conducted  18  nitrogen  and  mineral 
balances  in  simple  inflammation  of  the  lungs.  The  nitrogen  loss 
in  fever  was  found  to  be  dependent  on  the  intake ;  that  is,  the  great- 
er the  intake,  the  smaller  the  loss.  The  chlorine  retention  was 
much  higher  in  fever  than  when  the  temperature  was  normal.  The 
calcium  outgo  was  also  decreased  in  fever  while  the  phosphorus 
balances  were  usually  negative  and  seemed  to  follow  the  nitrogen. 

Sommerfeld  and  Caro  (1902)  studied  metabolism  in  three  child- 
ren who  were  convalescent  after  scarlet  fever,  with  a  diet  of  milk 
alone.      A  part  of  the  results  are  as  follows : 


518 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


AVERAGE   DAILY  NITROGEN  AND  PHOSPHORUS  BALANCES  WITH 

THREE   CHILDREN   CONVALESCENT   FROM   SCARLET   FEVER, 

ON  A  DIET  OF  PURE  MILK— Grams 


Case  No. 
and  age 

Milk 
taken 
C.C. 

Nitrogen 

Phosphorus 

Food 

Urine 

Feces 

Balance 

Food 

Urine 

Feces 

Balance 

I 
514  years 

II 
7  years 

III 
6  years 

3000 
3700 
2562.5 

16.395 
19.978 
13.688 

13.350 
15.423 
12.530 

0.965 
1.058 
0.535 

+2.080 
+3.497 
+0.623 

3.140 

3.875 

2.688 

0.663 
0.955 
1.205 

2.035 
2.205 
1.540 

+0.442 
+0.715 
+0.057 

Computed  from  authors'  data. 

The  periods  were  4  days  each  in  extent.  Cases  I  and  III  devel- 
oped nephritis  hemorrhagica  in  connection  with  scarlet  fever,  but 
recovered,  No.  I  quickly,  No.  Ill  slowly. 

The  above  experiments  were  conducted  when  the  patients  had 
been  free  from  fever  for  several  days,  but  remained  in  bed.  During 
the  balance  experiment  Case  III  showed  in  the  urine,  leucocytes,  red 
blood  corpuscles  and  kidney  epithelium.  The  phosphorus  retention 
with  these  children  was  in  Cases  I  and  II  quite  considerable ;  in  Case 
III,  however,  there  was  a  loss  of  phosphorus  with  an  intake  which 
would  normally  cause  retention  even  in  an  adult. 

Achard,  Laubry  and  Thomas  (1902)  studied  the  question  as  to 
whether  the  retention  of  chlorine,  which  occurs  in  the  organism  in 
so  marked  a  degree  in  the  course  of  certain  acute  diseases,  is  accom- 
panied by  a  retention  of  phosphates.  Daily  determinations  of 
chlorine  and  phosphorus  in  the  urine  were  made  in  the  different 
stages  of  a  number  of  diseases.  Then  injections  of  sodium  glycero- 
phosphate were  made,  and  the  urine  examined  again.  The  diseases 
studied  were  typhoid  fever  (7  cases),  pleurisy  (1  case),  pneumonia 
(5  cases),  rheumatism  (1  case),  pulmonary  tuberculosis  (1  case), 
and  asystole  (3  cases).  The  authors  found  no  parallelism  between 
the  chlorine  and  phosphates.  Increased  phosphorus  excretion  was 
a  frequent  occurrence  in  acute  cases  of  infectious  diseases.  In 
typhoid  fever  it  was  often  greatly  increased,  and  was  observed 
before  the  chlorine  crisis,  while  there  was  no  general  improvement, 
and  while  the  fever  continued.  Increased  phosphorus  excretion 
occurred  even  in  the  period  of  the  acme  of  certain  diseases,  as  was 
observed  in  typhoid  fever  and  infectious  icterus,  and  so  can  not  be 
looked  upon  as  a  forerunner  of  recovery.  This  increased  phospho- 
rus excretion  was  often  independent  of  the  diet.  The  injected 
glycerophosphate  was  usually  quickly  eliminated  in  the  urine. 


PHOSPHORUS  METABOLISM  519 

According  to  Gouraud  (1902),  the  phosphorus  elimination  is 
similar  in  pneumonia  and  typhoid  fever,  in  absolute  value  as  well  as 
in  relation  between  phosphates  and  alkalis.  The  urinary  phosphates 
are  diminished  in  amount  during  the  time  when  the  temperature  is 
at  its  highest  point,  and  also  as  long  as  that  temperature  lasts.  At 
the  beginning  of  convalescence  there  is  a  "phosphate  crisis"  which 
varies  in  intensity  according  to  the  disease  and  its  duration.  During 
the  fever  the  percent  of  phosphate  earths  in  the  total  phosphates 
falls  from  45  to  30,  and  sometimes  even  as  low  as  10.  The  phenom- 
enon is  reversed  at  the  moment  of  crisis,  which  affects  especially 
the  earth  phosphates,  and  the  percent  of  phosphate  earths  in  the 
total  phosphates  increases  to  50  or  60,  and  sometimes  to  70  or  80. 

Gouraud  (1903)  believes  that  in  fever  the  disintegration  of 
phosphorus-containing  compounds  is  markedly  increased,  but  that 
the  katabolic  processes  are  not  carried  to  the  final  liberation  of 
inorganic  phosphoric  acid,  the  result  being  such  an  accumulation  of 
organically  combined  phosphorus  as  induces  degenerative  histologi- 
cal alterations.  Urinary  phosphates  are  decreased,  the  proportion 
of  P  to  N  signifying  the  seriousness  of  the  malady.  In  convales- 
cence enzymatic  activities  become  normal  and  the  accumulated 
organic  phosphorus  is  katabolized  and  eliminated,  the  result  being  a 
phosphaturic  crisis.  Death  is  sometimes  preceded  by  such  a  rapid 
disintegration  of  tissues  rich  in  phosphorus  as  results  in  an  ante- 
mortem  phosphaturic  crisis  the  proportion  of  P  to  N  in  the  urine 
becoming  above  normal. 

Garratt  (1904)  has  made  an  extensive  study  of  metabolism  in 
fever.  He  is  of  the  opinion  that  apparent  retention  of  katabolized 
phosphorus  in  fever  is  due  to  the  escape  of  the  phosphorus-rich  pro- 
teins from  the  oxidation  which  overtakes  phosphorus-poor  proteins, 
and  that  increased  excretion  of  phosphorus  in  fever  should  be 
regarded  rather  as  a  result  of  the  action  of  the  toxin  on  tissues  rich 
in  phosphorus  than  of  the  influence  upon  these  tissues  of  the  pyrexia 
itself,  therefore  that  it  has  but  little  direct  connection  with  the 
febrile  state.      This  article  contains  51  references. 

According  to  Molyakov  (1912),  sodium  nucleate,  given  in  the 
first  stages  of  scarlet  fever,  causes  a  great  increase  in  the  number  of 
polynuclear  leucocytes. 

See  also  W.  Zuelzer  (1876)  and  S.  Weber  (1901). 

Summary.  From  these  notes  it  is  apparent  that  in  fevers  the 
amount  of  the  phosphorus  outgo  is  the  resultant  of  the  effects  of  a 
considerable  number  of  agencies  differing  in  the  nature  as  well  as 
the  degree  of  their  influence,  as  determined  by  the  cause  and  dura- 
tion of  the  fever,  and  the  state  of  nutrition  of  the  patient. 


520  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Among  those  conditions  making  for  increased  outgo  of  phospho- 
rus are  (1)  increased  tissue  destruction,  (2)  acidosis  and  (3)  chlor- 
ine retention;  among  those  tending  to  restrict  the  outgo  are  (1) 
decreased  intake,  and  (2)  retention  for  growth  of  white  blood  cor- 
puscles and  pathogenic  organisms.  In  the  post-febrile  stages  there 
are  unusual  tendencies  making  for  phosphorus  retention,  to  com- 
pensate for  previous  losses,  but  this  retention  is  often  masked  by  an 
increased  outgo  due  to  leukolysis,  excretion  of  metabolic  wastes 
previously  delayed  by  unfavorable  conditions,  and  excretion  of 
products  derived  from  the  pathogenic  organism.  There  seem, 
therefore,  to  be  no  invariable  rules  as  to  interrelationships  of  urin- 
ary constituents  in  fevers  generally. 

GLYCEROPHOSPHATE  THERAPY 

On  account  of  the  facts  that  lecithin  phosphorus  is  absorbed 
largely  in  the  condition  of  glycerophosphoric  acid,  and  that  lecithin 
is  exceedingly  expensive,  considerable  interest  has  attached  to  the 
therapeutic  use  of  salts  of  glycerophosphoric  acid. 

Pasqualis  (1894)  found  that  the  calcium  glycerophosphate 
increased  the  urinary  phosphorus  a  day  sooner  than  did  calcium 
phosphate,  but  after  examination  of  the  blood,  concluded  that  the 
calcium  phosphate  was  only  slightly  slower  in  getting  into  the  circu- 
lation. The  glycerophosphoric  acid  was  not  found  unchanged  in 
the  urine,  even  when  large  amounts  were  given  per  os,  or  even  by 
injection.  By  a  new  method  he  finds  abundant  glycerophosphoric 
acid  in  the  blood.  Pasqualis  states  that  in  every  organ  the  glycero- 
phosphoric acid  is  converted  into  glycerin  and  phosphoric  acid. 

Robin  (1895)  published  urine  analyses  from  several  cases  of 
a  number  of  diseases  under  treatment  with  sodium  glycerophosphate 
by  subcutaneous  injection.  The  author  discusses  at  length,  and  in 
detail,  various  improved  conditions  of  metabolism  said  to  be  due  to 
this  treatment.  In  view  of  the  fact  that  no  feces  analyses  were 
made  we  are  unable  to  judge  of  the  validity  of  the  claims. 

De  Stella  (1897)  conducted  injection  and  feeding  experiments 
with  sodium  glycerophosphate  on  two  rabbits  and  a  dog.  Urine 
analysis  shows  increased  excretion  of  sodium  chloride,  urea  and 
phosphorus. 

Bardet  (1900)  distinguishes  between  the  therapeutic  effects 
of  acid  and  neutral  sodium  and  calcium  glycerophosphates.  The 
acid  glycerophosphates  can  not  be  used  hypodermically.  Urinary 
excretion  of  phosphates  was  not  appreciably  increased  by  the  inges- 
tion of  acid  glycerophosphates,  even  when  given  in  large  doses.  The 


PHOSPHORUS  METABOLISM 


521 


acid  glycerophosphates  increase  the  titratable  acid  of  the  urine.  In 
large  doses,  15-25  gm.  per  day,  they  have  a  purgative  and  chologogue 
action. 

West  (1902)  reports  "immediate  and  pronounced"  results  from 
the  administration  of  sodium  and  calcium  glycerophosphates  in  8 
cases  of  neurasthenia. 

Street  (1902)  reports  that  glycerophosphates  are  most  useful 
in  all  cases  of  nervous  impairment  due  to  overwork,  or  excess  of  any 
kind,  in  premature  advance  of  age,  and  in  senility,  attended  by  gen- 
eral debility ;  the  benefits  from  their  protracted  use  are  said  to  be 
striking,  and  their  value  considerable  in  chronic  neuralgia,  in  sciat- 
ica (by  hypodermic  injections  along  the  nerve),  and  in  convalescence 
from  la  grippe,  and  acute  infectious  diseases.  Street  notes  that 
Magnin  of  Paris  asserts  that  in  diabetes  he  has  seen  the  sugar 
remarkably  diminished  by  glycerophosphates.  To  get  the  best 
results  it  is  said  to  be  necessary  to  administer  the  glycerophosphates 
from  three  to  six  months,  sometimes  longer,  though  occasional  inter- 
ruptions of  several  weeks  in  the  course  of  very  prolonged  adminis- 
tration are  beneficial. 

Fabiani  (1903)  reports  favorable  results  from  glycerophos- 
phate therapy  by  oral  or  subcutaneous  introducton  in  8  cases. 

J.  Smolenski  (1904)  investigated  the  therapeutic  effect  of 
glycerophosphates  with  19  infants,  one  to  eleven-and-a-half  months 
of  age,  and  15  children  of  one  to  five  years.  Both  sodium  and  calcium 
salts  were  used.  No  case  showed  unfavorable  results ;  many 
showed  improved  appetite,  and  gain  in  weight. 

Novi  (1904),  in  balance  experiments  on  himself,  studied  phos- 
phorus metabolism  as  affected  by  antirabes  treatment,  with  and 
without  accompanying  sodium  glycerophosphate,  taken  per  os  or 
hypodermically.      The  numerical  results  are  as  follows : 


AVERAGE  DAILY  PHOSPHORUS  BALANCES  WITH  A  MAN  AS 

AFFECTED  BY  ANTIRABES,  WITH  AND  WITHOUT  SODIUM 

GLYCEROPHOSPHATE— Grams 


Period 

Length 
in 
days 

Intake 
P2O5 

Urine 
P2O5 

Feces 
P2O5 

Bal- 
ance 
P2O5 

Percent  of  total 
P2O5  ingested 

Urine 

Feces- 

Total 

1 

4 

2.4485 

1.3345 

0.8023 

+0.3117 

54 

32 

86 

Normal 

2 

3 

2.2827 

1.3452 

0.9702 

-0.0327 

59 

42 

101 

Cure,  nonvirulent  material 

3 

5 

2.1346 

1.4573 

0.7119 

-0.0346 

68 

33 

101 

Cure,  virulent  material 

4 

5 

2.0041 

1.4150 

0.8190 

-0.2300 

70 

40 

110 

Cure,  virulent,  and  0.200  gm.  sodi- 
um glycerophosphate,  per  os. 

5 

1 

2.3387 

1.4858 

0.5889 

+0.2640 

63 

25 

88 

Cure,  virulent  material 

6 

3 

2.3161 

1.4177 

0.6581 

+0.2403 

61 

28 

89 

Cure,  virulent,  and  0. 20  or  0.30  gm. 
sodium  glycerophosphate  injected 

7 

1 

2.2335 

1.3319 

0.5788 

+0.3228 

59 

25 

84 

Cure,  virulent  material 

622 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


The  author  attributes  to  the  antirabes  treatment,  both  virulent 
and  nonvirulent,  a  tendency  to  increase  the  urinary  phosphorus, 
this  being  due,  Novi  thinks,  to  the  same  constituent  of  the  injected 
material,  which,  while  not  itself  toxic,  produces  a  leucocytosis  and  a 
coexistent  or  following  leucolysis. 

The  figures  show  that  the  administration  of  sodium  glycero- 
phosphate, either  hypodermically  or  per  os,  increased  the  feces  phos- 
phorus, and  increased  the  loss,  or  decreased  the  retention,  of  this 
element.  Ingestion  per  os  did  not  reduce  the  urinary  phosphorus, 
reckoned  as  percent  of  the  intake,  but  hypodermic  injection  may 
perhaps  have  done  so  to  a  slight  extent. 

Frey  (1906)  has  obtained  favorable  results  from  therapeutic  use 
of  a  glycerophosphate  preparation  containing  iron  and  bromine. 

It  would  seem,  therefore,  that  there  is  at  least  similarity  in  the 
action  of  glycerophosphates  and  lecithin,  though  the  probability  is, 
of  course,  that  the  glycerophosphates  are  less  effective  through  the 
lack  of  those  functions  or  superiorities  of  lecithin  which  depend  on 
its  partial  absorption  in  an  unchanged  condition. 

For  an  extended  review  on  glycerophosphate  therapy  see  Merck 
(1911). 

PHOSPHORUS  METABOLISM  IN  GOUT 

Gout  is  due  to  uric  acid  retention,  and  results  from  deranged 
nuclein  metabolism.  Observations  on  phosphorus  elimination  with 
nuclein-free  and  nuclein-containing  diets,  therefore,  have  been  used 
in  judging  of  the  progress  of  this  disease.  Purin  base  elimination 
3nay  be  even  more  significant  as  indicating  incomplete  formation  of 
'uric  acid  or  urea. 

Hans  Vogt  (1901)  conducted  nitrogen  and  phosphorus  balances 
with  a  man  suffering  from  gout,  for  the  purpose  of  studying  nuclein 
metabolism  in  this  condition.      Balance  data  are  below. 

AVERAGE   DAILY   NITROGEN  AND  PHOSPHORUS   BALANCES   WITH 

GOUT  PATIENT  AND  CONTROL  AS  AFFECTED  BY  INGESTION 

OF  PANCREAS— Grams 


Gout  patient 

Control 

Body 
weig-ht  of 

gout 
patient 

Kg. 

Body 
■weight 

of 
control 

Kg-. 

Periods 

N 
Intake 
Balance 

P 
Intake 
Balance 

N 
Intake 
Balance 

P 
Intake 
Balance 

Diet 

Fore-period 
6  days 

Main  period 
5  days 

After-period 
4  days 

14.58 
+1.16 

19.31 
+3.79 

14.58 
-0.33 

2.621 
-0.513 

4.861 
+1.016 

2.621 
-0.535 

14.58 
-1.76 

19.31 
+2.01 

14.58 
—2.38 

2.621 
-0.938 

4.861 
+0.865 

2.621 
-1.108 

68.2-68.4 

68.4-68.95 

68.9-68.5 

64.3-64.9 
64.9-64.9 
64.9-64.8 

White    bread,    egg's, 
sirloin  steak,  butter, 
cream,  beer 
Same  plus  calf's  pan- 
creas 

Same  as  first  period 

PHOSPHORUS  METABOLISM  523 

The  subject  suffered  from  acute  gout,  caused  by  beer  drinking, 
and  also  from  kidney  trouble.  For  comparison  a  second  subject 
was  included,  a  servant  ordinarily  healthy,  but  suffering  from 
nervous  dyspepsia  during  this  experiment. 

The  gout  patient  was  35  years  old  and  formerly  healthy.  On 
Apr.  26,  1900,  typical  symptoms  of  gout  appeared.  After  treatment 
with  sodium  salicylate  he  showed  improvement.  The  experiment 
lasted  from  May  8-May  23,  1900. 

Conclusions.  1.  There  was  a  nitrogen  loss  in  the  preliminary 
and  after  periods,  due  to  insufficient  intake,  but  a  retention  by  both 
subjects  during  the  gland-feeding  period. 

2.  The  nuclein  is  entirely  split  in  the  organism.  The  phos- 
phorus is  excreted  normally  but  the  nitrogen  is  retained.  This 
nitrogen  retention  is  not  due  to  a  disturbance  of  the  secretory  activ- 
ity of  the  kidneys,  since  the  phosphorus  outgo  is  normal. 

3.  Much  uric  acid  was  retained  by  the  gout  patient  in  compari- 
son with  the  control.  This  is  the  result  of  a  disturbance  of  metabo- 
lism, which  may  be  due  either  to  a  decrease  of  oxidation  or  splitting, 
or  to  synthetic  processes.  The  outgo  of  phosphorus  as  compared 
with  the  outgo  of  nitrogen,  and  therefore  of  uric  acid,  in  the  case  of 
the  gout  patient  is  very  small,  compared  to  the  control.  This  points 
to  a  tardiness  of  nuclein  metabolism  of  the  gout  patient.  Whether 
the  large  retention  of  nitrogen  is  due  also  to  this  is  not  to  be  conclud- 
ed from  these  results. 

Waldvogel  (1902)  reports  a  metabolism  study  at  the  time  of 
a  gout  attack,  in  which  the  urine  was  examined  as  to  its  amount, 
specific  gravity,  total  nitrogen,  uric  acid,  total  phosphorus  and 
disodium  phosphate.      In  conclusion  he  says,  in  part : 

"The  gouty  attack  may  be  thus  characterized:  The  nitrogen 
retention  between  attacks  shows  itself  to  be  a  slowing  up  of  the 
elimination  of  the  decomposition  products  of  nucleins,  the  values 
for  both  uric  acid  and  phosphoric  acid  being  small,  without  any 
recognizable  kidney  insufficiency 

"But  while  the  phosphate  shows  gradual  increase  for  a  long 
time  after  reduced  nuclein  metabolism,  the  elimination  of  uric  acid 
rises  sharply  from  the  first  day  of  the  attack,  and  it  is  also  increased 
in  the  blood  bringing  about  an  elevation  of  8  without  kidney  insuffi- 
ciency making  its  appearance.  The  uric  acid  apparently  rises  from 
that  being  produced  in  the  joints,  the  solution  of  it  in  the  alkaline 
blood  taking  place  so  rapidly  that  the  kidney  secretion  cannot 
immediately  make  up  for  the  subsequent  appearance  of  uric  acid  in 
the  blood. 


524 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


"Also  before  the  attack  the  relative  amount  of  P205  and  uric 
acid  in  the  urine  is  different;  on  the  second  day  before  the  attack 
the  amount  of  uric  acid  is  already  low,  that  of  phosphoric  acid  not. 
Not  till  the  day  before  the  attack  does  the  value  for  P205  fall, 
while  that  of  uric  acid  does  not  appear  further  reduced." 

The  data  are,  in  part,  as  follows : 

URIC  ACID  AND  PHOSPHORUS  ELIMINATION  IN  THE  URINE  OF  A 
GOUT  PATIENT  FOLLOWING  ACUTE  ATTACK 
Waldvogel  (1902)— Grams 


Day 

Uric  acid 

Total  P2O5 

Na2HP04 

1 

0.652 

0.58 

0.18 

Day  of  attack 

2 

0.769 

0.71 

0.14 

3 

0.770 

0.61 

0.19 

4 

0.824 

0.74 

0.24 

5 

0.742 

0.81 

0.34 

6 

0.679 

0.88 

0.34 

7 

0.758 

1.48 

0.70 

8 

0.723 

1.43 

0.60 

9 

0.620 

1.25 

0.67 

10 

0.768 

1.57 

0.64 

11 

0.615 

1.37 

0.41 

12 

0.540 

1.21 

0.51 

24 

0.644 

Two  days  before  a  second  attack 

25 

0.479 

1.20 

6.42 

Kaufmann  and  Mohr  (1902)  found  that  the  values  of  endogen- 
ous alloxur  nitrogen  and  the  ratio  of  uric  acid-nitrogen  to  base-nitro- 
gen both  fell  within  normal  limits  in  gouty  subjects.  They  give  the 
following  nitrogen  and  phosphorus  balances  for  5  gouty  persons  on 
nuclein-free  diet. 

NITROGEN  AND  PHOSPHORUS  BALANCES  WITH  GOUT  PATIENTS  ON 
A  NUCLEIN-FREE  DIET— Grams 


Days 

Calories 
intake 

Nitrogen 

Phosphorus  (P2O5) 

Case 
No. 

Intake 

Balance 

Intake 

Balance 

Case 

1 
2 
3 
4 
5 

4 
3 
4 
3 

8 

3200 

? 
2900 
3200 
2350 

12.5-13.9 

21.25 

15.65 

13.6-19.3 

15.1-16.7 

+1.55 
+4.72 
-2.56 
+0.24 
-1.55 

8.'45 

6.25 
6.35-7.35 
5.78-6.33 

+i.'i6 

-0.77 
+0.71 
+0.13 

Chronic  gout 

Chronic  gout,  "very  fat" 

Acute  attack 

Acute  attack 

Less  acute  attack 

Judging  from  the  values  and  the  signs  of  the  nitrogen  and  phos- 
phorus balances,  it  is  supposed  that  in  Case  2  there  was  a  building 
up,  and  in  Case  3  a  breaking  down  of  phosphorized  nitrogenous 
tissue,  while  in  Cases  4  and  5  it  appears  that  the  material  used  was 
non-phosphorized  protein. 


PHOSPHOKUS  METABOLISM  525 

These  authors  also  made  observations  on  the  absorption  and 
retention  of  the  nitrogen  and  phosphorus  of  thymus  when  added  to 
the  purin-free  diet,  and  in  one  case  of  chronic  gout  they  added  to 
the  diet  alternately  "dark"  (filet)  and  "light"  (veal  or  haddock) 
meat ;  the  dark  meat  gave  no  greater  increase  of  alloxur  bodies  than 
did  the  light  meat. 

Ciuffini  (1910)  compared  the  calcium,  magnesium  and  phospho- 
rus balances  on  a  normal  subject,  a  subject  with  chronic  gout,  and  a 
subject  having  gout  with  paroxysms.  Balances  were  noted  during 
4-day  periods  on  mixed  diet  and  on  the  same  without  meat  (consid- 
ered to  be  purin-free).  Urinary  phosphorus  elimination  appeared  to 
be  markedly  low  in  chronic  gout.  In  paroxysms  there  was  phospho- 
rus retention  before,  during  and  after  the  highest  point  of  the 
attack. 

Rotky  (1910)  studied  nuclein  metabolism  in  various  diseases 
of  human  subjects,  including  gout,  and  he  reports  purin  nitrogen 
observations  on  urine  and  feces,  and  P205  on  the  urine.  Uric  acid 
excretion  resulting  from  a  milk  diet  was  considered  to  be  of  endogen- 
ous origin.  Exogenous  excretion  was  studied  by  means  of  the 
addition  of  guanin,  hypoxanthin  or  thymus  to  the  diet.  Serious 
disturbance  of  ferment  activity  was  noted  in  gout,  the  food  purins 
not  being  converted  to  uric  acid,  and  in  acute  nephritis  there  was 
delay  of  excretion,  and  but  a  small  part  of  the  purin  nitrogen  was 
excreted  as  uric  acid,  while  urinary  purin  bases  showed  a  small 
increase.  But  slight  variation  was  noticed  in  the  values  of  phos- 
phoric acid. 

With  regard  to  the  etiology  of  gout  as  an  anomaly  of  purin 
metabolism  the  reader  is  referred  to  the  discussion  of  Brugsch  and 
Schittenhelm  (1910).  The  discussion  includes  normal  nuclein 
metabolism,  as  well  as  its  disturbances,  and  was  preceded  by 
exhaustive  investigations  on  certain  phases  of  the  subject  (Schitten- 
helm, 1907 ;  Brugsch  and  Schittenhelm,  1907a,  1907b,  1907c,  1907d, 
1907e,  1907f,  1908-9,  1910;  Brugsch,  1909).  The  reports  of  others 
are  also  extensively  quoted. 

According  to  these  authors  uric  acid,  originating  in  nucleins, 
becomes  so  abundant  in  the  blood  as  to  exceed  its  solubility  limits 
in  that  fluid,  and  hence  urate  is  deposited  in  the  tissues,  which 
deposit  causes  the  arthritis,  pain  and  fever  of  gout.  The  deposits 
are  found  to  be  mainly  monosodium  urate.  Brugsch  and  Schitten- 
helm distinguish  a  metabolic  gout  and  a  kidney  gout,  according  as 
the  uricaemia  is.  due  to  disturbances  in  the  purin  metabolism  or  in 
the  excretion.  Disturbance  of  the  kidneys  may  follow  ae  a  second- 
ary result  in  metabolism  gout. 


526  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

In  metabolism  gout  they  find  uric  acid  constantly  present  in  the 
blood,  even  on  purin-f ree  diet  (endogenous  uricaemia) ,  and  in  four- 
fifths  of  all  the  cases  which  they  observed  the  uric  acid  excretion, 
during  intervals  between  attacks,  was  subnormal  or  at  a  low  normal 
value,  and  the  base-N  excretion  was  within  normal  limits.  The 
ratio  of  uric  acid-N  to  purin  base-N  excretion  tends  toward  the  low- 
er limit  of  normal  variation.  At  the  time  of  attack,  and  immediate- 
ly afterward,  the  uric  acid  elimination  was  high.  The  accumulation 
of  exogenous  uric  acid  (exogenous  uricaemia)  is  not  considered  a 
retention  in  the  usual  sense,  but  a  temporary  increase  due  to  a 
depression  and  a  retardation  of  urea-formation,  from  the  purin 
bodies  fed,  and  of  the  decomposition  and  elimination  of  uric  acid. 
According  to  Brugsch  and  Schittenhelm  this  depression  and  retard- 
ation involve  the  whole  fermentative  system  of  uric  acid  formation 
and  decomposition  (especially  nuclease  and  purin  deamidase,  togeth- 
er with  the  uricolytic  ferment,  and  to  a  far  less  degree  the  xanthin 
oxidase).  See  Miller  and  Jones  (1909)  for  evidence  throwing  some 
doubt  on  this  point.)  This  weakening  of  the  ferment  activity  raises 
the  uricaemia  to  hyperuricaemia,  and  the  symptoms  of  gout  appear. 

Of  phosphorus  metabolism  during  gout  we  may  say  then  that 
in  acute  attacks  the  balances  may  be  positive  or  negative.  The 
phosphorus  excretion  falls  before  the  attack,  is  low  during  the 
attack,  and  rises  slowly  thereafter  during  a  considerable  period.  In 
chronic  gout  the  urinary  phosphorus  excretion  is  low. 

INFLUENCE  OF  HEMORRHAGE  ON  PHOSPHORUS  METABOLISM 

Hawk  and  Gies  (1904)  in  their  work  on  the  influence  of  exter- 
nal hemorrhage  on  dogs  included  determinations  of  phosphorus 
balance.  The  experiments  here  reported  were  on  two  dogs,  normal 
at  the  beginning  of  the  work,  and  subjected  to  anaesthesia,  opera- 
tion and  hemorrhage.  The  blood  was  let  from  the  saphena.  The 
diet  was  meat,  crackers  and  bone  ash.  As  a  result  of  the  many 
alterations  in  the  usual  course  of  metabolism  blood  letting  seems 
slightly  to  favor  phosphorus  retention.  The  excess  of  ingested 
over  excreted  phosphorus  was  least  in  the  anaesthesia  period  (Exp. 
1,  period  3)  and  greatest  in  the  anaesthesia-operation  period  (Exp.  1, 
period  4) .  Although  the  results  were  not  very  striking,  operation 
and  hemorrhage,  in  most  instances,  seemed  to  cause  slight  retention 
of  phosphorus;  etherization  favored  increased  elimination.  The 
anaesthetic  used  was  principally  ether,  with  a  very  little  chloroform 
at  the  beginning. 


PHOSPHORUS  METABOLISM 


527 


INFLUENCE  OF  EXTERNAL  HEMORRHAGE  ON  PHOSPHORUS 
METABOLISM  IN  THE  DOG— Daily  Balances— Grams 


Exp, 

and 

period 


1-1 

1-2 


1-3 
1-4 

1-5 

1-6 

1-7 


2-1 
2-2 
2-3 


Initial 
weight 

of 

animal 

Kilos 


16.96 
16.29 


15.81 
15.44 

15.18 

13.76 

13.35 


11.85 
11.40 
10.47 


Treatment 


Normal 

Anaesthesia.operation  and  1st 
hemorrhage,  2.93  percent,  body 

weight 

Anaesthesia  alone 

Anaesthesia    and    operation, 

(no  hemorrhage) 

2nd  hemorrhage,  3.22  percent, 

body  weight 

3rd  hemorrhage,  3.51  percent, 

body  weight 

4th  hemorrhage,  3.26  percent, 
body  weight 


Normal 

Anaesthesia  and  operation.... 
Anaesthesia,  operation  and 
hemorrhage,  3. 11  percent,  body 
weight 


Meat 


0.557 

0.557 
0.558 

0.558 

0.543 

0.527 

0.527 


0.440 


Food  phosphorus 


Crack- 
ers 


0.094 


0.070 


Lard 


0.026 


0.013 


Bone 

ash 


1.779 


1.423 


Total 


2.456 

2.456 
2.457 

2.457 

2.442 

2.426 

2.426 


1.950 


Phos- 
phorus 
balance 


Days 

in 
period 


+0.045 

+0.123 
+0.015 

+0.163 

+0.022 

+0.065 

-0.180 


-0.030 
-0.025 


-0.017 


HOOKWORM 

Vannini  (1900)  conducted  balance  experiments    with    human 

beings  under  treatment  for  anaemia  from  hookworm  (Anchylostoma). 

Nothing  characteristic  was  observed  in  the  phosphorus  metabolism. 


ICTERUS 

The  connection  of  icterus  (jaundice)  with  phosphorus  metabo- 
lism is  comparatively  slight  and  indirect.  Neither  the  overloading 
of  the  blood  with  bile,  nor  the  deflection  of  bile  from  the  intestine, 
nor  yet  the  injury  which  sometimes  occurs  to  the  liver  cells  bears 
directly  on  phosphorus  metabolism  in  important  ways. 

Through  under-nutrition  there  is  sometimes  an  increased 
katabolism  of  body  protein,  and  hence  an  increased  production  and 
secretion  of  endogenous  uric  acid,  but  only  to  such  extent  as  is  due 
to  the  increased  tissue  destruction.  The  increased  gastric  secretion 
of  hydrochloric  acid,  which  is  usually  present,  probably  has  a 
demonstrable  bearing  on  phosphorus  metabolism,  as  also  the  charac- 
teristic calcium  soap  stools. 

Phosphorus  metabolism  in  icterus  has  been  studied  by  Simnit- 
ski  and  Rodoslawow  (1902)  and  by  Schilling  (1901a).  Simnitski 
and  Rodoslawow  found  urinary  phosphorus  nearly  normal,  though 
often  increased,  with  relative  amounts  of  constituents  normal. 


528  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Schilling  notes  that  Szydlowski  has  stated  in  a  dissertation, 
"Beitrage  zur  Mikroskopie  der  Faces,"  that  in  icterus  the  feces 
contain  no  crystals  of  tricalcic  phosphate.  Schilling,  however,  finds 
that  although  in  this  disease  the  feces  do  not  usually  contain  many 
phosphate  crystals,  when  the  food  has  a  high  "phosphorus  content 
they  are  readily  seen.  He  finds  no  relation  between  the  fatty-acid 
content  of  the  feces  and  the  amount  of  phosphate  crystals  present. 

LECITHIN  THERAPY 

The  use  of  lecithin  as  a  drug  has  been,  in  large  part,  due  to  its 
prominence  as  a  constituent  of  the  brain,  and  to  the  established 
increase  of  formation  of  its  cleavage  products  in  degenerating  nerve 
tissue.  The  conception  of  administering  nutriment  to  the  brain, 
direct,  is  attractive,  and  nothing  is  more  natural  than  that  this  idea 
should  have  been  accepted  by  the  ignorant,  overworked  by  the 
humbug,  and  stoutly  denied  by  the  honest  radical.  The  extreme 
claims  of  many  advocates  of  the  use  of  lecithin  in  therapeutics  have 
tended  so  strongly  to  discredit  any  such  element  of  truth  as  they 
may  have  expressed  that  it  is  with  reluctance  that  we  attempt  to 
point  out  the  probable  facts. 

Approaching  the  task  with  the  assumption  of  the  sincerity  of 
the  average  man,  we  have  eliminated  some  of  the  obviously  unrelia- 
ble claims,  have  toned  down  many  of  the  remainder,  and  have 
sought  to  call  attention  especially  to  those  points  on  which  there  is 
general  agreement.  With  this  explanation  it  seems  unlikely  that 
the  following  reports  should  require  any  further  discount  than  is 
implied  in  the  suggestion  that  there  is  more  inclination  to  record 
and  to  publish  the  favorable  than  the  unfavorable  results,  and  we 
can  only  speculate  as  to  the  number  of  unfavorable  results  which 
have  been  suppressed. 

Baud  (1858a,  1858b)  reported  beneficial  effects,  in  cachexia, 
from  the  administration  of  phosphorized  fatty  matter  extracted 
from  the  medulla  oblongata  of  animals. 

B.  Danielewsky  (1895a,  1895b)  states  that  subcutaneous  injec- 
tion of  lecithin  in  dogs  increased  the  number  of  red  corpuscles  up  to 
800,000  to  1,000,000  above  normal,  and  also  increased  the  haemo- 
globin. This  improvement  occurred  in  a  few  days  after  the  injec- 
tion, and  lasted  for  some  time. 

Serono  (1897a,  1897b,  1902)  conducted  subcutaneous  injection 
experiments  with  specially  purified  lecithin  on  a  considerable  num- 
ber of  pathological  human  subjects.  There  resulted  gain  in  appe- 
tite, and  improvement  in  general  condition,  a  rapid  increase  of  red 


PHOSPHOKUS  METABOLISM  529 

blood  corpuscles,  and  usually  increased  nitrogen  elimination  in  the 
urine.  Lecithin  was  found  to  possess  a  tonic  effect  comparable  to 
arsenic,  but  acting  more  quickly.  Haemoglobin  increased  very 
slowly,  and  only  after  improvement  of  general  conditions. 

Serono  considers  it  better  to  administer  lecithin  hypodermically 
than  by  ingestion,  because  of  digestive  decomposition  in  the  alimen- 
tary tract,  and  would  use  egg  lecithin  rather  than  brain  lecithin, 
because  of  admixture  of  the  latter  with  toxic  substances,  in  doses 
of  20-30  centigrams  or  more  per  day. 

Magnani  (1898)  found  injections  of  lecithin  helpful  during 
treatment  of  the  eyes,  by  improving  the  condition  of  the  cornea, 
when  it  had  suffered  during  lowered  nutrition  of  the  whole 
organism. 

Muggia  (1898)  reports  nutritive  and  curative  value  of  lecithin 
and  egg  yolk  injected  in  cases  of  anaemia  and  infantile  athrepsia. 

Saint-Aubin  (1900)  conducted  hypodermic  injection  experi- 
ments on  five  patients,  with  a  solution  obtained  by  filtration  of  a 
mixture  of  vaseline  oil  and  egg  yolk,  after  heating  to  coagulate 
nucleins  and  albuminoids.  The  treatment  led  to  increase  of  weight 
and  of  urea  elimination,  due,  it  is  said,  to  stimulation  of  cellular 
multiplication,  and  in  consequence,  of  activity  of  the  elements. 
Phosphorus  and  uric  acid  outgo  were  also  increased. 

Morichau-Beauchant  (1901)  has  made  an  extended  study  of 
lecithin  therapy,  and  reviews  experimental  work  beginning  with 
that  of  Danielewsky  in  1895.  From  his  conclusions  we  condense 
the  following : 

Lecithin  administered  either  subcutaneously  or  through  the 
stomach  stimulates  the  appetite,  and  leads  to  rapid  gain  in  weight. 
It  stimulates  growth,  and  increases  the  number  of  red  blood  corpus^- 
cles  and  the  percent  of  haemoglobin. 

It  increases  urea,  total  nitrogen  elimination,  and  the  coefficient 
of  utilization  of  nitrogen,  and  diminishes  uric  acid. 

In  tuberculosis  lecithin  is  of  service  in  the  earlier  stages,  not 
as  a  specific,  but  as  an  auxiliary.  Similarly  it  may  be  of  value  in 
diabetes  through  improving  general  conditions.  It  also  gives  good 
results,  as  a  rule,  in  anaemias,  cachexias  and  convalescence.  In 
neurasthenia  it  is  especially  valuable. 

Lecithin  is  active  in  doses  of  0.2-0.3  gm.  per  os  and  0.05-0.10  gm. 
subcutaneously  each  day,  or  every  other  day.  Lecithin  is  not 
toxic  in  any  dose.  Subcutaneous  administration  is  indicated  in 
gastrointestinal  disorders. 


530  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Suzor  (1901)  treated  two  patients  by  subcutaneous  injeetion  of 
raw  egg  yolk  in  water  solution,  and  obtained  stimulation  and  im- 
provement of  general  condition. 

Iljin  (1901)  conducted  feeding  experiments  with  a  laboratory 
helper,  on  mixed  diets  of  normal  foods,  to  which  brain  and  egg  yolk 
were  added  as  carriers  of  organic  phosphorus.  From  this  work  we 
have  seen  only  Maly's  brief  abstract.  The  organic  phosphorus 
compounds  were  considered  to  have  promoted  nitrogen  retention. 

Zaky  (1901a,  1901b)  studied  the  effects  of  lecithin  ingestion,  in 
doses  of  0.3  gm.  per  day,  on  urinary  elimination.  In  experiments 
with  three  subjects  there  appeared  to  be  slight  increase  of  total 
nitrogen  and  urea,  with  decrease  of  uric  acid  and  phosphorus.  The 
changes  were  so  small  as  to  be  of  doubtful  significance  unless  estab- 
lished by  most  critical  experiment. 

Answering  Robin's  criticism  to  the  effect  that  egg  yolk 
increases  uric  acid  excretion  Zaky  says  that  it  is  the  nuclein  and 
not  the  lecithin  of  the  egg  which  increases  the  uric  acid  elimination. 

Aries  (1902),  on  the  strength  of  clinical  experience,  considers 
lecithin  a  valuable  remedy  in  all  cases  where  "denutrition"  is  to  be 
combatted,  especially  in  old  age.  He  speaks  of  it  as  a  bioplastic  and 
morphogenic  agent. 

Maillon  (1902)  made  a  clinical  and  physiological  study  of  leci- 
thin by  administration  of  0.3-0.8  gm.  doses  in  34  cases  of  tubercu- 
losis, chlorosis  and  nervous  diseases,  basing  conclusions  principally 
on  an  examination  of  the  urine.  He  found  the  amounts  of  phos- 
phoric and  glycerophosphoric  acids  in  the  urine  unchanged  by 
the  lecithin  treatment.  The  ingestion  of  lecithin,  however,  was  of 
benefit;  in  some  cases  it  improved  the  appetite;  in  chlorosis  it 
caused  a  constant  increase  in  body  weight,  but  in  tuberculosis  the 
increase  in  weight  was  irregular.  The  effect  on  the  urine,  if  there 
was  any  at  all,  was  indirect. 

Stassano  and  Billon  (1902a,  1902b,  1902c,  1902d)  conducted 
a  series  of  experiments  on  the  effects  of  lecithin  injection  on  the 
blood  elements  in  animals.  Lecithin  in  physiological  salt  solution 
injected  into  the  auricular  vein  of  rabbits  produced  hyperleucocy- 
tosis  lasting  for  several  days,  and  an  increase  in  the  ratio  of  mono- 
to  polynuclear  forms. 

Intraperitoneal  injection,  with  guinea  pigs,  caused  an  increase 
of  both  forms  of  leucocytes.  In  the  lecithin  exudate  the  ingestion 
of  the  polynuclear  forms  by  the  very  large  mononuclear  leucocytes 
proceeded  with  a  rapidity  suggesting  that  the  mononuclear  forms, 
especially,  had  taken  up  the  lecithin  injected,  and  nourished  them- 
selves therewith. 


PHOSPHORUS  METABOLISM  531 

Further  intravenous  injections,  with  rabbits,  led  to  a  progress- 
ive increase  of  red  blood  corpuscles  after  each  injection.  Tests 
showed  an  increased  vitality  of  these  cells,  as  implied  by  increase 
of  resistance  to  saline  solutions  of  various  strengths.  A  different 
reaction  of  the  red  corpuscles  to  stains,  implying  increased  acidity 
of  the  nuclear  chromatin,  was  noted. 

By  experiments  with  frogs  it  was  determined  that,  after  having 
grown  large  and  granular  at  the  expense  of  the  lecithin,  the  mono- 
nuclear leucocytes  left  the  blood  vessels,  and  spread  into  the  system. 
Endothelial  cells  in  their  turn  retained  lecithin,  becoming  crammed 
with  granulations,  and  their  nuclei  showing  marked  activity. 

Courtial  (1903)  showed  that  1-2  gm.  of  lecithin  taken  by 
healthy  people  produced  no  notable  differences  in  the  composition  of 
the  urine ;  no  increase  of  either  phosphorus  or  uric  acid. 

Fratta  (1904)  conducted  hypodermic  injection  experiments  on 
a  human  being  weighing  about  70  kg.  with  marrow,  sodium  hypo- 
phosphite,  glycerophosphates,  lecithin  and  cura  antirabica.  Glycero- 
phosphates and  lecithin,  injected  hypodermically  in  doses  of  0.20 
gm.  daily,  increased  urinary  nitrogen  elimination,  but  decreased  the 
urinary  phosphorus  excretion.  The  cura  antirabica  increased  both 
nitrogen  and  phosphorus  elimination. 

Heim  (1904)  describes  "bioson"  as  a  protein-iron-lecithin 
combination  and  recommends  it  as  a  blood  building  and  nerve 
strengthening  preparation.  The  iron  content  is  0.24  percent,  and 
the  lecithin  1.27  percent.  It  is,  therefore,  not  nearly  so  rich  in 
lecithin  as  many  common  foods. 

Migliaccio  (1904)  used  lecithin  injections  in  cases  of  anaemia, 
atrophy  and  rachitis, — all  infants.  The  results  were  rapid  and 
sometimes  notable  gain  in  weight,  notable  improvement  in  the 
blood,  and  in  gastrointestinal  functions.  The  urinary  phosphorus 
showed  increase  in  all  cases. 

Silberstein  (1904)  discusses  and  recommends  phosphorus 
treatment  with  "hemaprotagon"  tablets,  a  preparation  containing 
iron  from  the  blood,  and  unaltered  lecithin.  Cases  of  melancholia, 
tuberculosis  and  lues  are  discussed,  and  the  preparation  recommend- 
ed for  such  cases. 

W.  Koch  (1905a)  stated  that  for  the  clinical  administration  of 
lecithin  to  the  adult,  in  the  pure  form,  there  is  at  present  no  good 
scientific  basis.  Koch  based  this  view  on  the  fact  that,  in  the 
ordinary  diet,  one  consumes  vastly  more  lecithin  than  is  recom- 
mended by  the  promoters  of  the  clinical  application  of  lecithin.      He 


632  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

suggests,  however,  that  with  the  infant  the  case  may  be  slightly 
different  in  that  the  lecithin  content  of  cow's  milk  diluted  with 
water  may  be  reduced  to  a  figure  very  much  below  that  in  human 
milk. 

Snowman  (1905)  gives  clinical  reports  of  benefit  from  the  use 
of  sanatogen,  especially  improved  blood  conditions  in  chlorosis, 
chorea,  and  tuberculosis. 

For  suggestions  regarding  the  therapeutic  use  of  lecithin  activ- 
ated by  Rontgen  rays,  radium  rays,  and  by  other  means  see  Werner 
(1905). 

Von  Oefele  (1907)  found  "bioson"  to  be  utilized  in  a  normal  way 
during  health,  and  in  sickness  to  be  utilized  more  perfectly  than 
other  articles  of  diet. 

Errani  (1908)  treated  10  pathological  cases  by  lecithin  injec- 
tion. He  concluded  that  lecithin  promotes  and  accelerates  the  nor- 
mal  bioplastic  and  morphogenetic  processes,  improves  the  metabo- 
lism and  the  constitution  of  the  blood,  and,  in  consequence,  the 
general  condition  of  the  organism. 

Kleinertz  (1908)  reported  several  instances  of  improvement  in 
clinical  condition  after  the  use  of  "biocitin,"  a  lecithin  preparation 
of  Altmann,  containing  10.7  percent  of  lecithin. 

Sleeswyk  (1908)  found  that  the  bactericidal  action  of  certain 
lecithin  solutions  on  typhus  bacilli,  as  reported  by  Bassenge,  was 
due  to  their  acid  reaction,  caused  in  all  probability  by  impurities  in 
the  commercial  products  used. 

W.  Koch  (1909a),  in  discussing  phosphorus  compounds  as  brain 
foods,  calls  attention  to  the  facts  that,  in  comparison  with  other 
parts  of  the  body,  the  nervous  system  does  not  have  a  very  large 
amount  of  total  phosphorus,  though  it  has  of  lecithin;  that  the 
amount  is  not  materially  less  in  cases  of  dementia  praecox,  but  is 
reduced  in  general  paralysis ;  that  the  commercial  phosphorus  prep- 
arations used  as  drugs  (hypophosphites,  glycerophosphates,  phytin, 
lecithin)  are  much  less  effective  for  supplying  the  requirements  for 
growth  of  the  brain  than  are  phosphorus-rich  foods,  some  of  them 
not  being  taken  up  by  the  system,  the  amount  usually  recommended 
of  any  kind  giving  very  insignificant  addition  to  the  amount  of  phos- 
phorus taken  with  the  daily  food,  and  there  being  no  conclusive  evi- 
dence that  they  have  any  effect  on  the  growth  of  the  brain. 

J.  and  W.  Cronheim  (1910)  made  clinical  observations  on  the 
use  of  lecithin  with  18  cases,  all  of  them  undernourished,  and  most 
of  them  tuberculous.  The  authors  conclude  that  under  such  con- 
ditions lecithin  induces  remarkable  increase  of  appetite,  and  conse- 
quent gain  in  weight. 


PHOSPHORUS  METABOLISM  633 

Renshaw  and  Atkins  (1910)  studied  the  bactericidal  properties 
of  lecithin  and  of  choline  salts.  Such  a  property  of  lecithin  was 
shown  to  be  at  the  most  very  slight,  while  there  was  no  indication 
of  bactericidal  effect  of  the  choline  salts. 

Buslik  and  Goldhaber  (1911)  conducted  metabolism  experi- 
ments with  a  lecithin-albumin,  "glidine."  This  preparation  appeared 
superior  to  scraped  meat  as  to  digestibility,  absorbability  and  action 
on  intestinal  putrefaction ;  it  permitted  forced  protein  storage,  and 
had  a  favorable  effect  on  the  haemoglobin  content  of  the  blood  of 
anaemics ;  in  the  last  respect,  however,  "f erroglidine"  was  still  more 
efficient. 

Borschim  (1911)  applied  lecithin  to  the  back  of  a  rabbit  to 
determine  its  effect  on  iodine  absorption.  The  result  was  negative. 
Lecithin  has  no  such  effect. 

Milkowicz  (1911)  found  that  lecithin  and  cholesterol  in  vitro 
cause  a  great  increase  of  phagocytosis  of  Staphylococcus  and  tubercle 
bacilli. 

An  iodine-lecithin-protein  preparation  is  described  by  Isaac 
(1911)  and  C.  Neuberg  (1911)  under  the  name  "iodocitin." 

Bain  (1912)  obtained  from  the  oral  administration  of  lecithin 
to  rabbits  marked  increase  of  both  red  and  white  corpuscles  and  of 
haemoglobin,  and  the  same  results  from  subcutaneous  injection. 
The  increase  of  white  corpuscles  was  chiefly  in  the  mononuclear 
lymphocytes. 

Similar  experiments  were  conducted  with  phytin.  Introduced 
by  mouth,  the  improvement  in  blood  conditions  was  slight ;  introduc- 
tion by  injection  produced  marked  increase  in  red  corpuscles,  but 
less  increase  of  white  corpuscles  and  of  haemoglobin  than  was 
obtained  by  the  injection  of  lecithin. 

Bain  regards  lecithin  especially  as  a  metabolic  stimulus,  its 
effects  on  the  nervous  system  being  secondary  to  improvement  in 
general  state  of  nutrition. 

In  our  own  experience  we  have  found  that  egg  yolk,  presum- 
ably through  its  lecithin  content,  often  improves  the  digestion  of 
milk  by  infants,  but  also  that  infants  differ  widely  in  their  tolera- 
tion of  egg  yolk. 

Among  other  articles  on  lecithin  therapy  are  those  of  Coulombe 
(1901),  who  presents  a  historical  review  up  to  1901,  with  summary 
and  56  references;  Hartenberg  (1901),  Huchard  (1901),  Lancer- 
eaux  and  Paulesco  (1901),  Labbe  (1903),  Fiirst  (1903),  Grimm 
(1903),  H.  C.  Wood  Jr.  (1903),  Goliner  (1905),  G.  Landsberg 
(1906)  and  Berkley  (1908),  the  above  consisting  of  clinical  reports 


534  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

of  the  use  of  lecithin  in  a  great  variety  of  pathological  cases  char- 
acterized, in  general,  by  low  state  of  nutrition,  as  after  wasting 
diseases,  the  results  in  almost  all  cases  reported  being  marked 
improvement  of  general  condition. 

It  would  appear  from  these  clinical  reports  that,  in  states  of 
impoverishment,  lecithin  is  a  valuable  aid  in  the  restoration  of  the 
organism  to  normal  condition.  Claims  of  stimulation  of  growth 
under  normal  conditions  are  not  so  well  sustained,  and  are  difficult 
of  adjudication  by  reason  of  the  fact  that  the  normal  state  is  vari- 
able and  not  definitely  definable. 

The  benefit  derived  from  lecithin  treatment  is  usually  spoken 
of  as  improvement  of  the  general  state  of  nutrition,  especially  as 
evidenced  by  improved  appetite  and  digestion  (which  indeed  might 
be  accountable  for  most  of  the  rest),  increased  weight,  red  blood 
corpuscles,  haemoglobin,  and  white  blood  corpuscles,  especially  of 
the  mononuclear  form.  It  is  said  not  to  be  specific  in  disease,  but 
to  be  of  value  in  many  derangements, — anaemia,  cachexias,  conval- 
escence, neurasthenia,  etc.,  through  the  elevation  of  the  general 
plane  of  nutrition. 

As  to  the  relative  efficiency  of  lecithin  administered  as  a  drug 
and  taken  in  natural  foods  we  have  not  information  sufficient  to 
warrant  a  positive  statement.  If  lecithin,  taken  in  the  pure  form, 
actually  has  the  capacities  which  it  is  said  to  possess,  this  may  be 
due  to  the  more  extensive  absorption  in  an  unsplit  condition  which 
seems  probable  when  the  compound  is  administered  uncombined 
with  nutrients  requiring  digestive  cleavage. 

See  also  notes  on  lecithin  therapy  in  the  discussion  of  tubercu- 
losis. For  an  extended  review  on  lecithin  therapy  see  Merck  (1912). 

MALNUTRITION  OF  THE  BONES  OF  THE  DOMESTIC  ANIMALS 

The  facts  of  the  existence  of  malnutrition  of  the  bones  as  a 
disease  of  the  domestic  animals,  the  dependence  of  this  disorder 
primarily  on  deficiencies  in  the  mineral  elements  of  the  food,  as 
determined  by  its  nature  and  by  soil  and  climate,  and  the  ready 
response  of  this  ailment  to  improved  treatment,  either  by  change  of 
food,  use  of  fertilizers  on  the  pastures,  or  direct  feeding  of  bone- 
producing  salts,  have  been  demonstrated  many  times  over  in  a 
long  series  of  agricultural  investigations  in  which  there  is  such 
general  agreement  that  individual  statement  of  the  results  is  unnec- 
essary. The  following  notes  suggest  some  of  the  kinds  of  observa- 
tions to  be  found  in  the  literature. 


PHOSPHORUS  METABOLISM  585 

According  to  Moussu  and  Dollar  (1905)  osseous  cachexia  was 
known  in  Norway  as  early  as  1660. 

Koloff  (1866)  recognized  pregnancy,  lactation  and  digestive 
disorders  as  predisposing  conditions,  and  (1869)  doubled  the  calcium 
and  phosphorus  content  of  meadow  grass  by  the  use  of  fertilizers. 

E.  Voit  (1880),  experimenting  with  dogs  and  pigeons  on  lime- 
poor  and  other  diets,  produced  rachitic  conditions  by  withholding 
lime.^  Animals  of  the  larger  breeds  suffered  more  acutely  from  lime 
deficiency  than  those  of  the  smaller,  slower-growing  breeds.  The 
bones  were  decreased  in  phosphorus,  magnesium  and  iron  as  well  as 
calcium,  by  the  calcium  starvation. 

Nessler  (1873)  demonstrated  the  connection  of  this  disorder 
with  the  nature  of  the  underlying  rocks  from  which  the  soil  was 
derived.  Stilling  and  von  Mering  (1889)  produced  malnutrition  of 
the  bones  in  a  dog  by  use  of  a  lime-poor  diet.  Maier  (1894)  reports 
this  disorder  in  pigs  fed  on  low-calcium  foods.  Von  Seelhorst, 
Georgs  and  Fahrenholtz  (1900)  showed  that  the  moisture  of  the 
soil  affected  the  phosphorus  of  the  forage.  Phosphatic  slag  gave 
best  results  as  a  fertilizer,  through  increasing  the  phosphorus  of 
the  plants.  Lane  (1906)  discusses  this  bone  disease  in  army 
horses,  mules  and  donkeys  in  South  Africa.  Dibbelt  (1911)  found 
that  pups  in  utero  developed  normally  while  the  mother  lived  on  such 
a  low-calcium  diet  as  caused  puerperal  osteomalacia. 

Schenke  (1903)  quotes  Pott  and  Dammann  in  their  recom- 
mendations of  the  amounts  of  precipitated  calcium  phosphate  to 
feed  to  farm  animals.  Pott  advises  its  constant  use  with  young 
animals ;  swine  for  6  months,  colts  and  calves  for  the  first  year.        ' 

Dammann  advises  feeding  to  a  pregnant  and  milk-giving  sheep 
12-20  gm.,  a  sow  the  same,  a  mare  and  a  cow  30-50  gm.  Wherever 
there  is  fear  of  disturbance  of  normal  bone  formation,  foals  and 
calves  may  be  given  8-15  gm.  daily,  and  pigs  and  lambs  3-6  gm. 

For  young  fowls  Pott  advises  a  teaspoonful  of  bone  meal  for 
each  12-14,  for  hens  2-3  gm.  each. 

Under  normal  conditions  such  additions  are  not  necessary,  but 
with  food  poor  in  calcium  or  phosphorus  they  may  be  needed. 
Schenke  summarizes  a  large  number  of  investigations  on  the  use  of 
precipitated  calcium  phosphate  for  animals. 

Dobrowolsku  (1911)  has  found  that  phosphorized  cod-liver  oil 
is  fairly  efficacious  in  relieving  the  experimental  malnutrition  of 
the  bones  which  results  from  the  establishment  of  fistulae  in  the 
alimentary  tract. 


536  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

See  also  the  following  articles  having  to  do  with  malnutrition 
of  the  bones  in  domestic  animals:  Bopp  (1838),  Haubner  (1854, 
1867),  Nessler  (1866-71),  H.  Bauer  (1868),  Karmrodt  (1867),  Stoh- 
mann  (1869),  Tripier  (1874),  Haubner  (1875),  C.  Voit  (1877b),  J. 
Cohnheim  (1882),  Stutzer  (1888),  Kellner,  Kohler  and  Barnstein 
(1894,  1895),  Lafevre  (1894),  Winkler  (1891),  Bongartz  (1894a), 
Uhlich  (1895),  Robert  (1895),  Bonnetat  (1903),  Moussu  and  Dollar 
(1905),  Alquier  (1905-6),  Grandeau  (1905),  Lewite  (1905-8), 
Klimmer  and  Schmidt  (1906),  Ingle  (1907,  1908,  1909),  Scheunert, 
Schattke  and  Loetsch  (1911),  and  Neubauer  and  Hillkowitz  (1912). 

We  have  not  considered  infectious  osteoporosis  and  "big-head." 

MENTAL  AND  OTHER  NERVOUS  DISORDERS 

In  considering  the  influence  of  mental  diseases  on  the  phospho- 
rus metabolism  of  the  body  as  a  whole,  as  measured  by  the  excreta, 
we  should  bear  in  mind  the  facts  that  the  central  nervous  system 
is  not  highly  vascular,  and  not  the  seat  of  intense  metabolic  change, 
that  it  contains  but  a  very  small  part  of  the  total  phosphorus  of  the 
body,  and  that  violent  functional  disorders  are  accompanied  by  but 
slight  observable  change  in  its  chemical  composition.  There  is, 
therefore,  no  reason  to  anticipate  marked  alterations  in  the  total 
phosphorus  outgo  directly  related  to  changes  in  mental  condition. 

We  have  at  hand,  however,  certain  evidence  which  bears  on 
this  matter,  and  we  submit  the  same,  in  brief,  with  the  suggestions 
that  such  effects  of  mental  disorder  as  have  been  observed  on  the 
phosphorus  outgo  are  at  least  largely  due  to  the  indirect  influence  of' 
the  disease  on  general  metabolism,  especially  on  muscular  activity, 
and  on  secretion  and  absorption  in  the  alimentary  tract,  with  the 
result  of  altered  rates  and  paths  of  elimination ;  that  in  some  cases 
the  mental  disorder  may  be  the  effect  rather  than  the  cause  of 
a  metabolic  disturbance;  and,  further,  that  in  the  absence  of  feces 
analyses  the  urine  data  are  of  doubtful  value  in  this  relation. 

NERVE  DEGENERATION  AND  PHOSPHORUS  METABOLISM 

In  the  search  for  recognizable  products  of  nerve  tissue  katabo- 
lism,  which  might  serve  as  measures  of  this  process,  considerable 
attention  has  been  bestowed  upon  choline,  particularly  in  the 
cerebrospinal  fluid,  since  Noll,  Mott,  Halliburton,  Donath  and  others 
have  demonstrated  a  slow  autolytic  liberation  of  lecithin  and  choline 
in  degenerating  nerve  tissue.  It  has  been  suggested  that  choline, 
which  is  slightly  toxic,  and  the  highly  poisonous  neurine,  which 
differs  from  choline  by  one  molecule  of  water,  may  contribute  to  the 
production  of  the  intoxication  of  fatigue,  as  well  as  acute  mental 
and  nervous  disorders. 


PHOSPHORUS  METABOLISM 


537 


Coriat  (1904a)  found  that  choline  was  produced  by  autolysis  of 
nerve  tissue,  and  also  by  putrefactive  decomposition  of  lecithin  or 
tissues  rich  in  lecithin,  but  not  by  cleavage  by  pepsin  or  trypsin. 

Nesbitt  (1899b)  and  Hoesslin  (1906)  have  found  choline  and 
neurine  as  products  of  intestinal  putrefaction,  and  Kutscher  and 
Lohmann  (1906)  have  demonstrated  neurine  in  the  urine  of  human 
beings. 

According  to  Halliburton  (1905)  the  existence  of  choline  in  nor- 
mal cerebrospinal  fluid,  and  in  salt  water  extracts  of  fresh  nervous 
tissue,  implies  chemical  activity  in  living  nerve  substance,  the  more 
active  part,  the  gray  matter,  yieldng  the  greater  amount  of  choline. 

Halliburton  considers  that  the  cerebrospinal  fluid  functions  as 
the  lymph  of  the  brain,  and  that  the  increased  choline  content  of 
the  cerebrospinal  fluid  in  dementia  paralytica  is  directly  due  to  decom- 
position of  brain  substance.  In  harmony  with  this  idea  is  the  find- 
ing by  Donath  (1904)  of  a  coincident  increase  of  phosphoric  acid 
in  cerebrospinal  fluid  in  cases  of  degenerative  lesion,  the  phosphoric 
acid  also  being  considered  as  a  product  of  lecithin  decomposition. 


PHOSPHORIC  ACID  (P205)   CONTENT  OF  CEREBROSPINAL  FLUID  IN 
DISEASE  (Donath,  1904)  Grams 


Diagnoses 


Maximum 


Minimum 


Mean 


Anaemia 

Neurasthenia 

Epilepsy 

Hysteria 

Pulmonary  tuberculosis  with  melancholia 

Water  on  the  brain 

Sclerosis  multiplex 

Tumor  on  the  brain 

Tabes  dorsalis    

Progressive  paralysis 


0.0070 
0.0278 
0.0209 
0.0093 
0.0026 
0.0116 
0.0051 
0.0286 
0.0426 
0.0508 


0.0064 
0.0028 
0.0036 
0.0065 
0.0026 
0.0082 
0.0045 
0.0068 
0.0046 
0.0052 


0.0067 
0.0153 
0.0086 
0.0076 
0.0026 
0.0100 
0.0048 
0.0177 
0.0203 
0.0219 


These  figures  show  the  highest  phosphoric  acid  content  of  the 
cerebrospinal  fluid  in  those  diseases  which  are  accompanied  by 
rapid  degeneration  of  nerve  tissue. 

For  comparison  with  the  above  we  have  Mestrezat's  average 
figure  (1911)  for  the  percent  of  P205  in  20  normal  cerebrospinal 
fluids  of  0.003. 

Mott  and  Halliburton  (1901)  found  choline  in  the  cerebrospinal 
fluid  and  also  sometimes  in  the  blood  in  cases  of  severe  nervous 
lesion,  especially  in  paresis,  and  Donath  (1903b)  considers  choline  a 
factor  in  the  causation  of  epileptic  seizures,  though  he  did  not  find 
choline  in  the  cerebrospinal  fluid  in  functional  disorders  not  associ- 
ated with  tissue  destruction.      See  also  Masoin  (1904). 


538  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Mott  and  Halliburton  (1901)  studied  Wallerian  degeneration, 
both  chemically  and  microscopically.  In  a  number  of  cats  they 
severed  both  ischiadici.  The  animals  were  killed  after  different 
intervals  of  time  had  elapsed  after  this  operation.  The  normal 
blood  of  the  cat  contains  but  the  merest  trace  of  choline,  but  in 
three  or  four  days  after  the  operation  it  became  quite  evident,  at 
which  time  degeneration  of  the  nerve  was  recognizable  by  Marchi's 
reaction.  These  symptoms  were  most  pronounced  in  the  nerve 
which  had  been  severed  for  8  days.  Coincident  with  the  increase 
of  choline  in  the  blood  was  an  increase  in  the  moisture  content  of 
the  nerve  and  a  decrease  in  the  phosphorus  content  of  the  solid 
substance  of  the  nerve.  Regeneration  was  fairly  complete  after 
100-106  days,  the  first  sign  of  the  return  of  phosphorus  being  the 
beginning  of  the  response  to  the  myelin  reaction  on  the  60th  day. 
In  the  normal  nerve  the  phosphorus  content  is  a  little  over  1  per- 
cent; in  the  regenerated  nerve  a  little  less  than  1  percent.  The 
moisture  content  returns  to  normal  when  regeneration  begins. 

These  authors  state  that  excessive  degeneration  of  nerve  tissue, 
either  in  general  paralysis  of  the  insane,  or  in  other  diseases,  is 
accompanied  by  an  accumulation  of  choline  in  the  blood,  and  of  loss 
of  phosphorus  in  the  nerve  accompanying  the  change  of  lecithin  to 
non-phosphorized  fat. 

Noll  (1899)  also  conducted  similar  experiments  with  dogs  and 
horses. 

Mott  and  Barratt  (1899)  studied  the  chemistry  of  nerve  degen- 
eration by  analysis  of  the  spinal  cord  after  hemiplegia.  (See  table, 
p.  539.)      Their  conclusions  were  as  follows: 

"On  the  degenerated  side  of  the  cord  in  simple  hemiplegia  it 
was  found  that  (1)  a  breaking  up  of  the  phosphorized  fat  occurs; 
(2)  the  amount  of  lecithin  present  is  diminished;  (3)  the  amount 
of  fat  present  is  in  excess ;  (4)  the  amount  of  extractives  soluble  in 
ether  is  increased;  (5)  the  proteid  residue  diminishes  in  amount 
pari  passu  with  the  increase  in  extractives;  (6)  the  phosphorus  in 
the  residue  diminishes  at  a  still  greater  rate  than  the  residue  itself; 
(7)  the  percentage  of  phosphorus  in  the  half -cord  as  a  whole  is  dim- 
inished ;  (8)  the  ether  extract  has  an  appearance  of  butter  instead 
of  being  crystalline." 

Coriat  (1904b)  has  studied  the  cerebrospinal  fluid  in  different 
nervous  diseases,  29  cases  being  investigated.  Choline  was  found 
in  25  and  quantitatively  determined  in  21 ;  was  absent  in  alcoholic 
depressive  hallucinosis,  a  senile  paranoiac  condition,  melancholia 
and  senile  dementia.    It  was  invariably  present  in  general  paralysis, 


PHOSPHOEUS  METABOLISM 


539 


but  no  parallel  could  be  observed  between  the  amount  of  choline,  the 
weight  of  the  brain,  and  duration  of  the  disease.  The  largest 
amounts  of  choline  seemed  to  be  associated  with  the  highest  per- 
centages of  protein,  both  being  considered  as  measures  of  the  extent 
of  destruction  of  the  central  nervous  system.  Cholesterin  was 
always  absent.  See  also  Apelt  and  Schumm  (1908)  and  Coriat 
(1903),  (34  cases  discussed,  21  titles  and  references). 


LECITHIN  AND  PHOSPHORUS  DETERMINATIONS  ON  THE  TWO 

HALVES  OF  SPINAL  CORD  AFTER  LEFT  HEMIPLEGIA 

Mott  and  Barratt    (1899)— Percent 


Case 

Lecithin 

P  in  ether  extract 

P  in  residue 

P  in  half-cord 

Left  side 

Right  side 

Left  side 

Right  side 

Left  side 

Right  side 

Left  side 

Right  side 

I 
II 

18.9 
20.7 

22.2 
23.5 

1.72 
1.84 

2.14 
1.93 

0.92 
1.03 

0.97 
0.98 

1.19 

1.38 

1.38 
1.42 

Ziveri  (1909)  also  found  choline  in  the  cerebrospinal  fluid  in 
paralysis. 

Symmers  (1904-5a,  1904-5b)  reports  that  in  nervous  diseases 
of  the  degenerative  type  the  absolute  and  relative  amounts  of 
organic  phosphorus  of  the  urine  are  increased,  sometimes  to  an 
enormous  extent,  to  such  amounts  as  he  considers  could  not  be 
derived  directly  from  the  destruction  of  nervous  tissues,  but  rather 
from  increased  production  of  phosphorized  endogenous  metabolic 
compounds,  or  as  a  result  of  lessened  oxidation,  with  organic  phos- 
phorus compounds  as  end  products. 

W.  Koch  and  W.  H.  Goodson  (1906)  published  a  prelim- 
inary study  of  nerve  tissue  degeneration.  A  normal  human  brain 
was  compared  with  the  brain  of  a  paralytic,  and  a  healthy  spinal 
cord  of  a  dog  with  a  degenerated  cord.  (See  numerical  data  p.  130.) 
They  found  a  smaller  content  of  solids  in  the  degenerated  brain  than 
in  the  normal.  Nucleoproteins  were  increased,  according  to  the 
authors,  by  leucocytes,  proliferating  blood  vessel  elements  and  neu- 
roglia cells.  The  cord  of  the  dog,  which  was  allowed  to  degenerate  for 
19  days  after  cutting,  had  a  composition  like  that  in  the  degenera- 
tion of  general  paralysis.  The  proportions  of  constituents  remained 
unchanged,  but  the  absolute  amounts  were  reduced.  The  authors 
state  in  conclusion,  "Nervous  tissue  more  than  any  other  tissue, 
both  in  pathological  and  in  experimental  degeneration,  tends  to  keep 
its  relative  composition  constant,  which  observation  is  in  harmony 
with  the  results  obtained  in  starvation." 


540  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

W.  Koch  and  Mann  (1909)  published  analyses  of  brains  of  nor- 
mal human  beings  and  also  of  brains  of  four  cases  each  of  dementia 
praecox  and  general  paralysis.  (See  p.  132.)  The  variations  in 
the  diseased  brains  were  within  the  range  of  variation  in  the  normal 
brains. 

The  present  state  of  knowledge  of  the  chemistry  of  the  brain 
and  of  methods  for  its  study  do  not  begin  to  answer  the  require- 
ments of  such  an  investigation. 

Pighini  (1911)  tested  for  esterase  the  blood  serum  in  various 
nervous  diseases  such  as  epilepsy,  idiocy  from  contusion  of  the 
brain,  dementia  praecox,  alcoholism,  etc.,  by  the  speed  of  reaction 
against  neutral  sodium  monobutyrate,  as  shown  by  developing 
acidity;  and  compared  these  observations  with  others  on  normal 
serum.  In  most  of  these  cases  the  action  was  slower  than  normal, 
although  quite  evident.  Similar  tests  with  a  lecithin  emulsion  in 
sodium  chloride  solution  showed  the  presence  in  all  these  cases  of  a 
lipase  which  would  split  lecithin,  but  only  in  the  presence  of  a  small 
amount  of  MnS04.  It  reached  its  maximum  activity  within  the 
first  hour. 

The  optical  method  (Pighini,  1910)  was  used  for  testing  the 
serum  for  nuclease.  In  cases  of  alcoholism  and  epilepsy  there  seems 
to  be  less  activity  of  the  nuclease  of  the  serum  than  is  normal,  dur- 
ing the  interval  between  attacks ;  while  in  acute  maniac  depressive 
insanity,  and  in  the  period  following  epileptic  attacks,  this  activity 
is  increased. 

Pighini  and  Nizzi  (1912)  report  that  neither  in  normal  condi- 
tions nor  in  dementia  praecox,  epilepsy  or  progressive  paralysis  is 
there  a  specific  esterase  or  lecithase  in  the  cerebrospinal  fluid. 

PHOSPHOEUS  METABOLISM  IN  EPILEPSY 

Lepine  and  Jacquin  (1879)  made  a  study  of  the  proportions  of 
phosphorus  and  nitrogen  in  the  urine  of  10  hospital  patients  and  a 
number  of  dogs.  In  the  case  of  certain  epileptics,  the  relation  of 
phosphorus  to  nitrogen  is  much  lower  than  normal  during  the  inter- 
vals between  attacks.  In  the  case  of  the  same  patients  the  relative 
amount  rises  remarkably  immediately  after  the  attack.  Sometimes, 
without  there  being  an  attack,  but  only  a  threat  of  an  attack,  the 
relative  amount  rises,  and  this  rise  may  be  due  to  the  alkaline 
earth  phosphate. 

Lailler(1884)  reached  similar  conclusions,  to  the  effect  that 
at  the  time  of  the  crisis  or  immediately  thereafter,  the  urine  con- 
tains an  increased  amount  of  phosphorus  and  a  small  amount  of 


PHOSPHORUS  METABOLISM  541 

urea,  but  when  the  crises  succeed  each  other  rapidly  both  phospho- 
rus and  urea  are  increased,  while  in  the  intervals  between  crises  the 
urine  is  normal. 

Mairet  (1884c)  investigated  metabolism  in  various  mental  dis- 
orders. In  epilepsy  he  found  during  attacks  an  increased  urinary 
excretion  of  nitrogen  and  phosphorus. 

Loewe  (1910)  also  found  an  increased  excretion  of  phosphorus 
in  the  urine  of  the  day  of  the  attack,  both  organic  and  inorganic 
phosphorus  sharing  in  the  increase,  the  latter,  however,  more  than 
the  former. 

W.  Koch  (1904)  published  a  brain  analysis  of  an  epileptic  who 
had  died  in  an  attack.  The  analyses  are  submitted  as  provisional, 
and  especially  as  illustrative  of  the  methods  of  analysis. 

COMPOSITION  OF  HUMAN  BRAIN  (EPILEPTIC) 
Koch  (1904)  Percent 

Corpus  Callosum  Cortex  (Prefrontal) 

Water 67.97  84.15 

Simple  proteids 3.20  (by  difference) 5.00  (by  difference) 

Nucleoproteids 3.70 3.00 

Neurokeratin 2.70  (Chittenden) 0. 40  (Chittenden) 

Extractives 1.51 1.58 

Inorganic  salts 0.82  0.87 

Lecithins 5.19  3.14 

Kephalin  and  myelin 3.49  0.74 

Amido  lecithins trace  trace 

Phrenosin  and  kerasin 4.57  1.55 

Cerebrin  acids trace  none 

Cholesterin „    4.86 0.70 

Sulphur  compound 1. 40  1. 45 

99.41  102.58 

Loewe  (1911)  determined  total  and  organic  phosphorus  in  the 
urine  of  33  subjects  for  1-12  days  after  attack  of  various  psychoses 
and  neuroses.  There  was  noted  an  increased  organic  phosphorus 
elimination  after  epileptic  and  some  other  attacks,  and  a  probable 
increase  in  certain  phases  of  paralysis  and  delirium  tremens,  though 
no  increase  in  a  number  of  other  diseases  in  the  psychiatric  realm, 
as  for  instance  in  kakatonia.  A.  Bornstein  (1911)  found  also  a 
slight  increase  in  the  serum  lecithin  in  epileptics.  This  increase 
of  the  lecithin  content  of  the  blood  serum  is  regarded  as  indicative 
of  increased  katabolism  of  brain  lipoids. 

PHOSPHORUS  METABOLISM  IN  PARALYSIS 

Ewald  (1883)  determined  phosphorus  in  the  urine  in  8  cases  of 
paralysis  agitans,  chorea  and  senile  tremor  in  periods  of  2-33  days, 
also  in  four  controls  in  periods  of  12-23  days.  The  urinary  phos- 
phorus excretion  was  not  in  any  way  characteristic. 

Gurther  (1883),  in  somewhat  extended  studies,  reached  the 
same  conclusion  with  cases  of  hystereoepilepsy  and  paralysis 
agitans. 


542  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Robert  and  Parisot  (1906)  studied  4  cases  of  paralysis  agitans 
with  trembling  and  one  without.  In  the  former  cases  they  thought 
that  the  urinary  phosphorus  excretion  was  subnormal,  but  the  basis 
for  their  conclusion  is  slight  or  of  doubtful  value. 

Barratt  (1899),  studying  the  water  and  phosphorus  in  the 
central  nervous  system  in  general  paralysis  of  the  insane,  and  other 
conditions,  found  that  there  was  a  decrease  in  the  phosphorus  of  the 
hemispheres  of  the  brain  associated  with  evidence  of  nerve-degen- 
eration and  of  chromatolysis ;  and  with  increase  in  percent  of  water, 
but  not  proportional  to  any  one  of  these  conditions,  or  to  the  age  of 
the  patient,  or  the  wasting  of  the  hemispheres.  A  similar  condi- 
tion was  found  in  the  spinal  cord. 

Halliburton  (1905)  says  that  in  general  paralysis  the  cerebro- 
spinal fluid  is  richer  than  normal  in  solids,  especially  in  proteins,  the 
most  abundant  of  which  being  nucleoprotein ;  and  much  richer  also 
in  choline,  both  nucleoprotein  and  choline  being  considered  as 
products  of  degeneration  of  nerve  substance.  See  also  Mott  and 
Halliburton  (1901)  and  Halliburton  (1901b). 

Glikin  (1909a)  estimated  the  lecithin  of  the  marrow  of  the  long 
bones  of  diseased  persons.  He  found  in  five  cases  of  dementia 
paralytica  and  in  one  of  tabes  (ages  33-43)  no  lecithin  or  only  a  trace, 
while  in  five  others  (ages  30-43)  he  found  1.195-4.21  percent.  Where 
lecithin  was  absent  phosphorus  and  iron  were  also  absent.  Glikin 
gives  the  following  figures  for  the  lecithin  content  of  the  fat  of  bone 
marrow  of  normal  men. 

Age  34   3.30  percent  lecithin 

Age  56  2.02  percent  lecithin 

Age  61   2.21  percent  lecithin 

Age  70   2.33  percent  lecithin 

Age  70   2.76  percent  lecithin 

Age  88  1.83  percent  lecithin 

A.  Bornstein  (1909)  found  an  increased  lecithin  content  of  the 
blood  serum  in  paralysis.  Lecithin  was  determined  by  the  Glikin 
chloroform  method.  He  gives  as  the  normal  content  of  lecithin  in 
blood  serum  2.0-2.4  percent;  in  progressive  paralysis  he  found  2.7- 
3.5  percent,  and  in  taboparalysis  2.9  percent. 

In  a  later  publication  (1911)  Bornstein  reports  11  cases  of 
paralysis  in  which  the  lecithin  content  of  the  blood  serum  was  deter- 
mined. Two  were  considered  as  within  the  normal  range  of  varia- 
tion, these  being  1.86  and  2.38  percent,  while  the  remaining  9  cases 
ranged  from  3.07-4.60  percent  lecithin  in  the  serum. 


PHOSPHORUS  METABOLISM 


543 


PHOSPHORUS  METABOLISM  IN  INSANITY 

Mairet  (1884c),  in  studying  phosphorus  metabolism  in  mania, 
divided  the  observations  into  four  periods  covering  different  por- 
tions of  the  attack  of  the  disease,  as  follows :  agitation,  depression, 
remission  and  convalescence.  The  results  of  this  investigation  are 
indicated  by  the  accompanying  table. 

URINARY  EXCRETION  IN  MANIA 


Periods 

Nitrogen 

Phosphorus 

Total 

Alkaline  earth 

Alkali 

Increased 
Diminished 

Increased 

Increased 
Diminished 

Increased 

Diminished 

" 

" 

In  melancholia  there  was  an  increase  in  the  alkaline  earth  phos- 
phates, and  a  decrease  of  the  alkali  phosphates  of  the  urine. 

Lailler  (1884)  reached  similar  conclusions;  that  in  acute  delir- 
ium or  mania  there  is  a  marked  excess  of  urea  and  of  phosphorus 
in  the  urine,  in  mania  with  excitement  a  slight  excess  of  phospho- 
rus only,  while  in  simple  mania,  or  in  melancholia  without  agitation, 
the  urine  is  normal.  In  acute  melancholia,  or  with  excitement, 
there  is  a  notable  increase  in  the  urea  and  a  slight  increase  in  the 
phosphorus  of  the  urine. 

Modica  and  Audenino  (1901)  found  in  10  cases  of  insanity, 
caused  by  immoral  practices,  a  reduction  in  the  alkali  phosphates 
of  the  urine,  and  in  certain  cases  also  the  total  phosphorus.  In  5 
healthy  guinea  pigs  and  2  healthy  dogs,  whose  frontal  lobes  had 
been  removed,  the  alkaline  earth  phosphates  of  the  urine  decreased, 
and  finally  disappeared,  while  the  total  phosphorus  and  the  nitrogen 
also  diminished. 

Folin  and  Shaffer  (1902)  studied  the  metabolic  accompani- 
ments of  a  48-hour  periodicity  of  general  nervous  disturbance 
diagnosed  as  maniac  depressive  insanity.  On  the  nervous  days 
there  was  an  increased  amount  of  phosphorus,  and  an  increased  pro- 
portion of  nitrogen  and  sulphur  to  phosphorus  in  the  urine.  The 
authors  concluded,  from  the  constancy  of  the  percentage  of  alkali 
phosphates  on  four  consecutive  days,  that  the  increase  of  phospho- 
rus elimination  on  the  nervous  days  was  not  due  to  katabolism  of 
bone  tissue. 

As  explaining  the  above  phenomenon  the  authors  suggest  that 
"there  exists  in  this  patient  on  every  second  day  a  condition  some- 
what analogous  to  diabetes,  in  virtue  of  which  the  system,  or  some 
part  of  it,  is  unable  to  assimilate  (organize)  a  part  of  the  phosphate 


544  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

absorbed  from  the  digestive  tract.  The  non-assimilated  phosphate 
is  eliminated  on  the  same  day,  and  the  total  amount  of  phosphate 
eliminated  on  the  'nervous'  days  is  therefore  greater  than  the 
amount  eliminated  by  a  normal  person  absorbing  the  same  amount 
of  phosphoric  acid  from  the  digestive  tract.  On  the  alternating 
days,  on  the  contrary,  a  correspondingly  less  amount  of  phosphate 
than  the  normal  is  eliminated,  because  on  those  days  the  system 
repairs  the  loss  sustained  on  the  preceding  days."  The  authors 
suggest  further  that  it  is  the  nervous  tissues  which  are  subject  to 
the  abnormal  periodicity  of  ability  to  assimilate  phosphates  which 
produces  the  above  described  phenomenon.  In  view  of  the  small 
amount  (about  12  gm.  P205)  of  the  total  phosphorus  of  the  central 
nervous  system,  however,  it  is  unlikely  that  the  very  considerable 
variations  (about  0.66  gm.  P205)  in  the  daily  urinary  phosphorus 
excretion  could  depend  directly  on  the  state  of  nutrition  of  the 
nervous  system.  This  investigation  included  no  feces  phosphorus 
estimations. 

At  a  later  date  Folin,  Shaffer  and  Hill  (1904),  after  a  careful 
study  of  the  urines  of  insane  patients  and  numerous  controls, 
reached  the  following  conclusions: 

"From  a  constructive,  positive  point  of  view  it  must  be  admitted 
that  they  teach  very  little  that  is  tangible  concerning  mental  dis- 
eases, except  for  the  strong  suggestion  which  they  contain  that  in 
general  paralysis  we  have  a  disease  which  may  be  associated,  at 
one  stage  or  another,  with  some  demonstrable  metabolism  disorder. 

"From  the  destructive,  negative  or  critical  point  of  view,  it  is 
believed  that  the  data  given  prove  the  untrustworthiness  of  all 
those  metabolism  experiments,  old  and  new,  which  report  a  'char- 
acteristic' increase  or  diminution  of  any  of  the  urinary  constituents 
included  in  this  research  as  associated  with  any  particular  one  of 
the  ordinary  mental  disorders. 

"It  is  not  claimed  that  such  abnormal  metabolism  may  not 
exist,  but  simply  that  the  experiments  recorded  in  the  literature 
are  insufficient  to  demonstrate  the  fact." 

Nizzi  (1912)  reports  that  the  elimination  of  nitrogen  and  phos- 
phorus is  increased  in  the  initial  stage  of  maniac-depressive  insan- 
ity and  decreased  in  the  chronic  stage. 

E.  Mendel  (1872)  reported  urine  analyses  from  110  patients 
suffering  from  various  types  of  insanity  and  other  nervous  diseases, 
and  of  healthy  persons ;  also  the  same  from  a  dog  and  rabbit  with 
the  brain  pierced  by  a  -needle.  In  the  light  of  our  present  under- 
standing, however,  the  results  do  not  seem  significant. 


PHOSPHORUS  METABOLISM 


545 


ADMINISTRATION  OP  PHOSPHORUS  COMPOUNDS  AND  OTHER  DRUGS  IN  MENTAL 
AND  NERVOUS  DISEASES 

Pfeiffer  and  Scholz  (1899)  made  a  general  metabolism  and  res- 
piration study  of  senility  and  paralysis  agitans  as  affected  by 
thyreoidin.*      Figures  on  phosphorus  balance  are  as  follows : 

AVERAGE   DAILY  PHOSPHORUS   BALANCES   IN   SENILITY   AND   IN 
PARALYSIS  AGITANS  AS  AFFECTED  BY  THYREOIDIN— Grams 


Exp.  No. 

and 

Condition  and 

Treatment 

Intake 

Urine 

Feces 

Balance 

duration 

age  of  subject 

P2O5 

P2O5 

P2O5 

P2O5 

in  days 

X    la- 6 

Par.  agitans,  57 

Without  thyreoidin 

3.3155 

2.3893 

3.1798 

-2.2374 

lb-5 

57 

With 

3.3593 

2.8518 

2.8703 

—2.3628 

2a-4 

57 

Without          " 

3.9479 

2.2340 

3.4550 

—1.7411 

2b-3 

57 

With 

4.3216 

2.4538 

3.4550 

—1.5872 

3a-4 

58 

Without 

2.7716 

2.0570 

2.4057 

—1.6911 

3b-3 

58 

With 

2.4491 

2.2669 

2.5385 

—2.3563 

5a— 6 

Healthy       76 

Without          " 

2.0906 

1.2095 

3.7702 

—2.8891 

5b-5 

76 

With 

2.2473 

1.8921 

2.3958 

-2.0406 

6a-4 

81 

Without 

2.0906 

1.3567 

2.5346 

-1.8007 

6b-5 

81 

With 

2.2655 

1.7317 

2.4873 

-1.9535 

These  results  show  both  classes  of  subjects  to  be  indifferent  to 
thyreoidin,  and  that  with  paralysis  agitans  there  is  a  marked 
increase  of  phosphorus  outgo  which,  on  the  diet  used,  appeared  in 
the  urine.  The  authors  regard  this  high  phosphorus  outgo  as  due 
more  especially  to  senility  than  to  paralysis  agitans. 

Gilbert  (1901)  reports  general  improvement  in  neurasthenics 
from  the  use  of  lecithin,  either  in  pills,  or  in  subcutaneous  injec- 
tions. 

Kaufmann  (1902)  studied  excessive  protein  ingestion  in  sub- 
acute neurasthenia,  with  a  man  22  years  old,  on  various  simple  diets, 
to  which  were  added,  in  certain  periods,  the  whites  of  30-39  eggs 
per  day.  This  excessive  protein  ingestion  caused  a  reduction  in 
the  excretion  of  phosphorus  by  both  urine  and  feces. 

Martinet  (1903)  writes  that  phosphate  therapy  is  beneficial 
both  in  accidental  and  in  long-standing  psychasthenia,  but  in 
psychatoxies  with  excitement  phosphate  therapy  aggravates  the 
pathological  state.  He  bases  his  conclusions  on  clinical  observa- 
tions and  examination  of  the  urine  after  administration  of  officinal 
phosphoric  acid,  with  sodium  acid  phosphate  added  to  decrease  the 
acid  taste. 

Donath  (1903a)  treated  progressive  paralysis,  as  well  as  toxic 
and  infectious  psychoses,  9  cases  in  all,  by  injection  with  a  salt  solu- 
tion composed  as  follows :  K2S04  0.25  gm.,  KC1  1.0  gm.,  NaCl  6.75 
gm.,  K2C03  0.40  gm.,  Na2HP04+12H20  3.10  gm.,  and  distilled  water 

*  Thyreoidin  is  the  same  as  thyroidin,  an  iodine-free,  crystallizable  compound  from  the 
thyroid;  an  amorphous  iodine-  and  phosphorus-containing  compound  from  this  gland  is  known 
variously  as  thyrein,  thyreiodin,  thyroiodin,  thyroiodinin  and  iodothyrin. 


546  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1000  gm.  This  solution  was  nearly  isotonic  with  the  blood.  The 
treatment  acted  as  a  stimulus  to  the  body  functions  generally,  as 
indicated  by  improved  heart  action  and  appetite,  increased  diuresis, 
and  particularly  as  a  tonic  to  the  nervous  system  as  shown  by 
improved  handwriting,  freshening  of  defective  memory,  increased 
accuracy  of  speech  and  singing,  and  ability  to  solve  problems,  etc. 

Gordon  (1906)  reported  clinical  observations  on  56  cases  of 
nervous  disease  under  treatment  with  phosphorus-containing  drugs, 
with  and  without  other  drugs.  Of  the  56  cases  28  were  multiple 
syphilitic  affections  of  the  nervous  system,  5  were  cases  of  tabes,  14 
were  neurasthenics,  2  melancholia,  3  cases  of  obcessions,  and  4  of 
cerebral  softening.  Phosphorus-containing  compounds  had  a  bene- 
ficial effect  on  the  asthenia,  and  apparently  also  improved  the  special 
symptoms  of  the  disease  itself.  Glycerophosphates  gave  the  best 
results. 

J.  Hoppe  (1907b)  has  studied  the  effects  of  adding  thyroid  tab- 
lets to  the  food  in  treatment  of  three  kinds  of  idiocy,  namely 
myxoedema,  cretinism,  and  "mongoloid"  idiocy.  In  myxoedema 
there  was  a  marked  increase  in  the  apparent  absorption  of  calcium 
and  phosphorus,  and  a  noticeable  growth  of  the  bones.  In  cretinism 
there  was  but  slight  improvement  in  the  absorption  of  calcium  and 
phosphorus ;  and  in  mongoloid  idiocy  none  at  all.  As  a  result  of  a 
vegetable  diet  and  rectal  feeding  with  5  gm.  Na2HP04  daily,  the 
patient  with  myxoedema  showed  improved  calcium  absorption  and  a 
simultaneous  increase  in  the  phosphorus  outgo  in  the  urine.  See 
alsoTuczek  (1884). 

Peritz  (1908,  1908-9a)  found  an  apparent  increase  of  lecithin 
in  the  feces  in  lues,  tabes  and  paralysis ;  and  also  that  lecithin 
injection  would  reduce  the  increased  lecithin  elimination  in  these 
diseases.  A  large  number  of  estimations  of  the  lecithin  in  the  blood 
serum  in  the  above-mentioned,  and  in  other  diseases,  were  presented 
(1908-9b).  The  results  were  characterized  by  much  variability. 
Lack  of  normal  controls  leaves  uncertain  the  significance  of  these 
observations  by  Peritz.  In  a  later  paper  Peritz  (1910)  shows  that 
the  lecithin  of  the  blood  serum  responds  readily,  by  marked  increase, 
when  lecithin  is  administered  per  os. 

Taghamuro  (1908)  administered  lecithin  by  injection  in  dorsal 
tabes.  There  resulted  improvement  of  digestion,  appetite  and 
general  condition,  as  well  as  increase  of  red  blood  corpuscles  and 
haemoglobin. 

Nerking  (1909)  made  an  extensive  study  of  the  effects  of  leci- 
thin on  narcosis.  With  considerable  numbers  of  rats,  dogs  and 
rabbits,  with  the  usual  use  of  controls,  he  treated  them  with  anaes- 


PHOSPHORUS  METABOLISM  547 

thetics  as  follows :  ether,  chloroform,  morphine,  scopolamin,  ureth- 
an,  urethan-chlorhydrate,  novokain-suprarenin,  tropakain  and 
stovain,  usually  as  subcutaneous,  intraperitoneal  or  intralumbar 
injections,  and  followed  these  treatments  with  injections  of  1-10  per- 
cent solutions  of  purified  lecithin.  In  all  cases  the  lecithin  injection  is 
said  to  have  produced  a  favorable  effect  on  narcosis  (1)  in  short- 
ening the  period,  (2)  in  the  alleviation  of  unfavorable  after-effects 
and  (3)  in  causing  a  rapid  return  to  normal  condition. 

Donath  (1909)  experimented  with  2-percent  sodium  nucleate, 
containing  an  equal  quantity  of  sodium  chloride,  by  subcutaneous 
injection  in  21  cases  of  general  paralysis,  in  amounts  of  50-100  c.c. 
at  intervals  of  5-7  days.  The  injections  caused  a  rise  of  tempera- 
ture, in  some  cases  to  40.5°  (average  38.5°),  and  an  increase  of 
leucocytes  to  61000  (mean  23000).  Out  of  21  cases  10  were  suf- 
ficiently improved  to  return  to  ordinary  duties  (the  oldest  case  was 
of  two  years'  standing) ;  in  5  cases  there  was  improvement,  while  6 
remained  unimproved. 

Klieneberger  (1911)  and  Lowenstein  (1911)  report  negative 
or  unfavorable  results  from  the  use  of  sodium  nucleinate,  by  injec- 
tion, in  progressive  paralysis. 

For  the  successful  use  of  a  lecithin  preparation  in  migraine  see 
Schottin  (1911). 

Leubuscher  (1913)  gave  elemental  phosphorus  in  10000  parts 
of  oil  in  9  severe  old  cases  of  epilepsy.  Seizures  were  reduced  by 
30  or  40  percent  in  3  cases,  "about  65  percent  in  two  others ;  in  3 
others  the  improvement  was  less  striking,  while  with  one  patient 
there  was  no  improvement.  The  greatest  benefit  was  obtained  in 
the  more  severe  cases.  Phosphorus  was  administered  for  15 
months  with  6  cases. 

NEPHRITIS 

Fleischer  (1881)  found  in  chronic  interstitial  nephritis  a 
decreased  excretion  of  phosphorus,  and  a  parallelism  between  nitro- 
gen and  phosphorus  elimination  by  the  kidneys.  Sodium  phosphate 
ingestion  led  to  quantitative  excretion  of  the  phosphorus  within  24- 
48  hours  in  a  normal  subject,  but  in  nephritis  produced  little  or  no 
increased  phosphorus  excretion — a  difference  not  noted  with  respect 
to  potassium  bromide  and  sodium  salicylate.  In  a  case  of  lighter 
acute  nephritis  there  was  a  marked  lowering  of  urea  and  phospho- 
rus in  the  urine.  . 

VanAckeren  (1890-91)  and  Kornblum  (1892)  found  neither  a 
parallelism  of  urea  and  phosphorus  elimination  nor  a  consistent  low- 
ering of  phosphorus  elimination. 


548  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Von  Moraczewski  (1896)  showed  that  in  parenchymatous  neph- 
ritis the  urine  contains  less  chlorine  and  more  phosphorus,  especially 
calcium  and  magnesium  phosphate,  than  normal.  In  interstitial 
nephritis,  however,  there  was  no  increase  of  urinary  phosphorus; 
in  fact  a  decrease  of  calcium  and  magnesium  phosphates.  Uric 
acid  was  found  increased  in  parenchymatous  nephritis  and  normal 
in  the  interstitial  variety. 

Mohr  (1903)  published  food  and  urine  analyses  from  four 
nephritic  patients.  The  data  show  plainly  that  the  phosphorus 
outgo  is  normally  affected  by  variations  in  the  intake. 

Von  Koziczkowsky  (1903-04)  found  in  nephritis  that  the  outgo 
of  phosphorus  varied  normally  with  the  intake,  and  that  the  phos- 
phorus outgo  was  indirectly  proportional  to  the  sodium  chloride 
retention. 

Kovesi  and  Roth-Schulz  (1904)  found  in  nephritis  that  the 
proportion  of  the  total  phosphorus  of  the  food,  which  appeared  in 
the  feces,  varied  widely  without  ascertainable  relation  to  food  or 
pathological  conditions.  In  their  cases  they  found  almost  constant 
phosphorus  retention,  reaching  very  high  figures  in  cases  of  rapidly 
developing  oedema ;  but  they  noted  no  relation  between  phosphorus 
elimination  and  the  form  or  symptoms  of  the  disease,  or  the  nitro- 
gen or  sodium  chloride  of  the  urine,  or  the  total  of  dissolved  mole- 
cules in  the  same.  These  authors  consider  it  desirable  to  restrict 
the  absorption  of  phosphates  in  nephritis,  and  they  note  von  Noor- 
den's  suggestion  that  calcium  carbonate  be  given  with  the  usual 
milk  diet  in  the  accomplishment  of  this  purpose. 

Von  Noorden  (1907,  II,  p.  467)  mentions  a  number  of  investi- 
gations, in  addition  to  those  which  we  have  seen,  in  which  the 
capacity  of  the  kidneys  to  eliminate  phosphorus  was  not  lowered, 
and  also  several  investigations  showing  that  the  observation  of 
von  Koziczkowsky  of  the  reciprocal  relation  of  phosphates  to  sodium 
chloride  was  not  the  rule;  hence,  we  are  unable  to  say  that  there 
is  any  invariable  character  of  the  urinary  phosphorus  elimination 
in  nephritis,  though  a  retention  of  phosphorus,  apparently  an 
expression  of  a  lowered  capacity  of  excretion,  is  very  common.  The 
significance  of  this  variation  is  not  known. 

Erben  (1903,  1905)  found  low  values  for  phosphorus  in  the 
blood  in  subchronic  nephritis,  in  chronic  parenchymatous  nephritis, 
and  also  in  cases  of  secondary  shrunken  kidneys. 

J.  Miiller  and  H.  Reinbach  (1913)  studied  digestive  lipaemia  in 
a  case  of  subacute  nephritis.  Lecithin  was  present  in  the  serum 
to  the  extent  of  0.688  percent. 


PHOSPHORUS  METABOLISM  549 

NUCLEIN  THERAPY 

This  subject  is  discussed  in  connection  with  normal  nuclein 
metabolism ;  see  p.  256-258. 

OEDEMA 
L.  E.  Meyer  (1905)  studied  the  effect  of  variations  in  the  water, 
sodium  chloride  and  phosphorus  content  of  the  food  of  a  nursing 
infant  suffering  from  idiopathic  oedema.  The  addition  of  200  c.c. 
of  distilled  water  per  day  to  the  food  was  without  marked  influence 
on  mineral  metabolism.  Where  sodium  chloride  was  fed  in  vary- 
ing quantities  the  storage  varied  directly  with  the  intake;  and 
where  sodium  phosphate  was  added,  the  sodium  chloride  intake 
remaining  constant,  there  was  marked  increase  in  the  storage 
of  both  phosphorus  and  sodium  chloride,  and  a  decrease  to  more 
nearly  the  earlier  retention  figures  when  the  phosphate  was  with- 
drawn. Meyer  was  of  the  opinion  that  the  above-mentioned  stor- 
age was  pathological  in  character,  in  that  the  outgo  was  not  nor- 
mally responsive  to  the  intake ;  that  the  cause  was  a  functional  dis- 
turbance of  the  epithelial  cells  of  the  uriniferous  tubules,  resulting 
in  an  overloading  of  the  blood  with  salts,  and  therefore  in  oedema; 
and  further,  that  the  treatment  for  idiopathic  oedema  should  be 
the  decrease  of  the  salt  content  of  the  food  to  the  lowest  possible 
amount. 

EFFECTS  OF  MISCELLANEOUS  ORGANIC  PHOSPHORUS-CONTAINING 

COMPOUNDS  AND  PREPARATIONS  ON  PHOSPHORUS 

METABOLISM 

TETBA-ETHYL-PHOSPHONITTM  IODIDE 

Lindemann  (1898)  studied  the  pharmacology  of  tetra-ethyl- 
phosphonium  iodide  by  injection  experiments  with  frogs,  rabbits, 
guinea  pigs  and  dogs.  The  action  of  this  substance  is  character- 
ized by  loss  of  coordination,  and  general  paralysis,  which  soon 
extends  to  the  respiratory  center  and  then  to  the  heart,  which  is 
invariably  found  engorged  and  in  diastole.  The  lungs  are  strong- 
ly hyperaemic  and  oedematous,  while  in  chronic  intoxication  there 
are  characteristic  changes  in  the  liver  and  kidneys,  consisting  in 
hydropic,  albuminotic  degeneration  of  the  protoplasm.  A  full 
statement  of  details  is  given.  This  substance  is  excreted  in  the 
urine  together  with  a  compound  giving  an  insoluble  precipitate  with 
barium,  and  a  volatile  base,  which,  with  sulphuric  acid,  forms  a  non- 
volatile salt.  The  only  marked  pathological  change  in  the  urine 
was  a  decided  haemoglobinuria,  appearing  in  the  last  stages  of 
chronic  intoxication.  It  is  therefore  shown  that  neither  the  lower 
oxidative  stages  of  phosphorus,  nor  phosphine,  nor  the  organic 
phosphorus  compounds  have  the  action  of  elemental  phosphorus. 


650  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

FEESAN 

Kornauth  and  Czadek  (1900)  experimented  with  fersan, 
a  beef -blood  preparation  containing  much  iron  and  phosphorus  in  a 
mixture  of  organic  combinations.  With  this  preparation  two 
experiments  were  conducted,  one  on  geese  and  another  on  a  man. 

Two  geese  were  fed  240  gm.  corn  each  per  day.  In  addition 
one  received  pellets  of  fersan.  In  two  7-day  periods  the  one  receiv- 
ing corn  alone  gained  70  and  230  gm.  in  live  weight,  while  the  one 
receiving  the  fersan  with  its  corn  gained  195  and  235  gm.  during 
the  same  periods. 

In  four  balance  experiments  of  6-15  days  duration  with  a  man, 
two  periods  with  fersan  and  two  without,  there  was  no  evidence 
that  this  preparation  could  improve  a  mixed  diet  containing  an 
abundance  of  meat,  though  the  authors  state  that  it  was  almost 
completely  absorbed  and  assimilated. 

EOBOEAT 

Laves  (1900)  studied  the  nutritive  value  of  prepared  proteins, 
roborat  (prepared  from  grain;  contains  lecithin),  tropon  (from 
flesh  and  blood),  aleuronate  (from  wheat),  and  plasmon  (from 
milk).  Experiments  of  short  duration  were  conducted  on  the 
author  and  on  a  dog,  with  artificial  digestion  tests.  Urine  analyses 
furnished  the  basis  for  conclusions.    The  results  are  not  significant. 

OAEBOHYDEATE-PHOSPHOEIC  ACID  ESTEES 

Euler  (1912b)  investigated  the  disposition  by  animals  of  the 
carbohydrate-phosphoric  acid  ester  synthesized  by  yeast.  His  con- 
clusions are  in  part  as  follows: 

This  ester  is  split  by  an  intestinal  enzyme  and  by  Bacterium  coli; 
also  by  an  enzyme  contained  in  the  kidney  of  the  horse.  The  cal- 
cium salt  of  this  ester  is  broken  down  and  the  inorganic  phosphate 
excreted  as  such.  The  calcium  salt  of  glycerophosphoric  acid  ester 
is  likewise  broken  down  by  Bacterium  coli.  The  calcium  salt  of  the 
phosphoric  acid  ester  when  fed  to  dogs  goes  over  largely  into  the 
urine  as  inorganic  phosphate. 

From  a  study  of  the  literature  Euler  concludes  that  organic 
phosphorus  in  the  food  is  absorbed  both  split  and  unsplit.  He  sug- 
gests that  "it  is  especially  to  be  emphasized  that  the  physiological 
effects  of  phosphates  are  to  be  sought  in  their  remarkably  strong 
influence  on  the  vital  enzyme  reactions." 

Euler  and  Funke  (1912)  report  an  experiment  with  the  carbo- 
hydrate-phosphoric acid  ester  formed  by  phosphatese,  the  compound 
being  fed  to  a  rabbit.  At  least  three-fourths  of  the  ester  under- 
went cleavage. 

For  discussions  of  nuclein,  lecithin  and  phytin  therapy  see  pages 
256,  528  and  315  respectively. 


PHOSPHORUS  METABOLISM  551 

PROTYLIN 

Protylin  is  a  phosphorus-containing  preparation  obtained  by 
the  dialysis  of  egg-albumin  after  protracted  treatment  with  ortho- 
phosphoric  acid.  It  is  a  white,  tasteless  powder,  insoluble  in  water, 
and  contains  2.7  percent  of  phosphorus. 

Gnezda  (1903)  reported  clinical  observations  on  11  cases  of 
cachexia  with  carcinoma,  hysteria,  anaemia,  nervous  dyspepsia,  etc., 
under  treatment  with  protylin,  which  is  said  to  have  proven  itself  a 
valuable  tonic,  as  evidenced  by  increase  of  appetite,  body  weight  and 
haemoglobin,  and  improved  psychic  conditions. 

Siegmann  (1904)  published  similar  observations  on  32  cases, 
as  also  did  Marian  von  Bilgorajski  (1904)  on  10  cases,  the  latter 
recommending  especially  the  iron  and  bromine  compounds.  See 
also  Dorn  (1904).      Laumonier  (1905)  submits  other  clinical  data. 

Pouchet  and  Chevalier  (1905)  conducted  injection  experiments 
with  a  protylin  solution,  38  gm.  protylin  and  2  gm.  soda  per  liter  of 
water,  with  dogs.  After  injection  for  2  hrs.  35  minutes  into  a 
chloralized  dog  a  sharp  decrease  in  blood  pressure  occurred,  and  an 
accelerated  and  subsequently  retarded  cardiac  action.  Afterward 
the  blood  pressure  slowly  became  normal,  if  the  injections  were 
continued  more  feebly,  the  cardiac  action  being  accelerated  simul- 
taneously. 

In  the  case  of  a  dog  with  section  of  the  pneumogastric  nerves 
there  took  place  a  more  marked  acceleration  of  cardiac  action,  the 
blood  pressure  decreased  less ;  on  the  other  hand  a  lessening  of  the 
cardiac  energy  followed,  which  caused  the  heart  to  fail  to  become 
completely  filled.  These  authors  consider  that  the  action  of  this 
phosphorus  preparation  on  the  circulation  accounts  to  a  great 
extent  for  its  influence  on  the  general  nutrition. 

Timpano  (1906)  reports  increase  of  appetite  and  of  strength, 
reviving  of  psychic  activity,  and  improvement  of  general  state  of 
nutrition  from  the  feeding  of  protylin  to  18  cases,  and  Gallenga 
(1906)  made  similar  observations  on  22  cases. 

R.  O.  Neumann  (1906)  showed  that  protylin,  when  used  in 
varying  amounts  to  replace  equivalent  amounts  of  cheese  protein  in 
a  mixed  ration,  does  not  increase  either  nitrogen  or  phosphorus 
retention.  When  added  to  a  full  diet,  however,  its  nutritive  value 
was  apparent,  and  both  nitrogen  and  phosphorus  storage  were 
increased. 

Fjodoroff  (1907)  determined  that  the  ingestion  of  lecithin, 
nucleic  acid  and  phytin  often  causes  a  slight  increase  of  free  hydro- 
chloric acid    in    the    stomach,    sufficient    in    cases    of    chronic 


552  OHIO  EXPEKIMENT  STATION:  TECHNICAL  BUL.  5 

hyperacidity  to  cause  heart-burn,  and  to  constitute  a  contraindica- 
tion of  their  use.  In  such  cases  protylin,  even  in  5-gram  doses,  is 
well  tolerated. 

See  also  Maestro  (1905a),  Wechsler  (1905)  and  Laguesse 
(1905). 

OSTEOMALACIA 

Osteomalacia  in  human  beings  is  a  disease  of  entirely  unknown 
cause,  one  of  the  prominent  symptoms  of  which  is  an  absorption  of 
the  salts  of  bone,  and  their  replacement  by  tissues  not  having  the 
normal  content  of  bone  salts.  It  is  not  due  primarily  to  lack  of 
phosphorus  or  calcium  compounds  in  the  food,  nor  to  rapid  growth, 
or  pregnancy,  or  lactation,  or  senility,  though  naturally  these  may 
all  be  contributory  or  accentuating  conditions ;  and  it  is  not  curable 
by  the  use  of  foods  which  are  rich  in  bone-forming  constituents. 

The  literature  of  this  disease  is  very  voluminous,  and  of  it  we 
have  seen  but  a  small  part.  There  is  little  variety  in  the  experi- 
mental findings,  and  our  materials  are  perhaps  typical  of  the  whole 
body  of  facts  as  published. 

LACTIC  ACID  IN  OSTEOMALACIA 

An  early  idea  regarding  human  osteomalacia  was  that  the  bone 
salts  were  removed  by  an  excess  of  acids  in  the  system,  and  lactic 
acid  seems  occasionally,  though  not  as  a  rule,  to  have  been  found  in 
the  blood,  urine  and  bones. 

C.  Schmidt  (1847)  reported  a  case  of  bone  disease  involving  all  of 
the  bones  of  one  leg,  resulting  in  the  absorption  of  nearly  the  whole 
of  the  mineral  substance,  the  periosteum  remaining  intact.  Analysis 
showed  the  soft  inner  portion  of  the  bone  to  contain  much  lactic 
acid;  but  Virchow  (1852)  discusses  a  fatal  case  of  puerperal  osteo- 
malacia in  which  the  bones  had  become  filled  with  a  soft  jelly-like 
substance  of  alkaline  reaction. 

O.  Weber  (1867)  found  lactic  acid  in  osteomalacic  bones,  and 
Steiner  (1869)  made  an  anatomical  study  of  senile  osteomalacic 
bones,  and  concluded  that  the  disease  was  due  to  removal  of  the 
lime  salts  by  lactic  acid. 

Note:  Osteomalacia  of  the  domestic  animals,  which  is  also  known  as  malnutrition  of 
the  bones,  halisteresis  of  the  bones,  fragility  of  the  bones,  and  osseous  cachexia,  differs  from 
human  osteomalacia.  This  malady  is  without  doubt  due  principally  to  deficiency  of  the  food  in 
bone-forming  constituents,  though  perhaps  also  in  part  to  a  lack  of  correct  proportion  between 
the  mineral  elements,  bone-forming  or  otherwise,  contained  in  the  food.  This  condition  is 
readily  curable,  simply  by  the  removal  of  the  cause,  though  in  the  course  of  time  affected 
animals  may  become  more  or  less  unresponsive  to  improved  conditions  of  diet.  The  principal 
predisposing  conditions  are  pregnancy,  lactation,  growth,  starvation,  and  unhygienic  surround- 
ings. The  use  of  the  term  osteomalacia  for  this  simple  malnutrition  of  the  bones  is  confusing 
and  should  be  abandoned. 


PHOSPHORUS  METABOLISM  553 

Mors  and  Muck  (1869)  found  much  lactic  acid  in  the  urine  of 
one  case  of  osteomalacia,  a  little  in  the  urine  of  another,  and  none 
in  the  urine  of  a  third  case.  They  consider  excessive  production  of 
acid  to  be  the  cause  of  osteomalacia.  In  one  case,  which  was  cured 
by  the  use  of  a  nourishing  diet,  cod-liver  oil,  and  a  salt  mixture  of 
calcium  carbonate,  calcium  phosphate  and  ferric  hydrate,  the  lactic 
acid  disappeared  from  the  urine  as  the  bones  began  to  harden. 

See  also  von  Jaksch ;  Ueber  die  Alkalescenz  des  Blutes;  Zeitschr. 
f.  klin.  Med.,  13,  355. 

Heitzmann  (1873)  administered  lactic  acid  subcutaneously, 
and  with  the  food,  to  7  cats,  5  dogs,  2  rabbits  and  1  squirrel,  the 
food  being  low  in  lime ;  and  the  experiments  were  continued  for  sev- 
eral months,  in  one  case  at  least,  13  months,  followed  by  examination 
of  the  bones. 

After  two  weeks'  administration,  by  either  method,  the  car- 
nivora  showed  rachitic  symptoms,  swelling  of  the  epiphyses  of  the 
long  bones  and  of  the  union  of  the  costal  cartilage  with  the  ribs. 
The  swellings  mentioned  increased  visibly  by  the  4th  and  5th  weeks ; 
the  long  bones  became  crooked,  and  the  microscopic  examination 
of  the  epiphyses  agreed  perfectly  with  that  of  rachitic  children. 

After  4  to  5  months'  feeding  of  the  acid  the  long  bones  became 
as  flexible  as  fish  bones.  The  microscopic  structure  of  the  bones 
after  4  to  11  months'  feeding  of  lactic  acid  was  analogous  to  that  of 
people  who  had  died  from  osteomalacia. 

With  the  three  herbivora  the  case  was  different.  The  walls 
of  the  long  bones  of  the  rabbits  became  very  thin,  but  remained 
brittle;  while  with  the  squirrel,  which  was  fed  for  13  months,  the 
walls  of  the  bones  became  very  thin  and  also  flexible. 

Unfortunately  in  these  experiments  there  were  present  two  pos- 
sible demineralizing  agencies,  and  it  is  impossible  to  say  whether 
the  results  were  due  to  calcium  starvation  alone,  or  in  part  to  the 
effects  of  the  acid. 

Vogt  (1875)  injected  into  the  tibiae  of  living  rabbits,  five 
weeks  old,  2  drops  of  pure  lactic  acid.  The  rabbits  were  killed  5 
months  afterward.  The  periosteum  was  rather  easily  separable 
from  the  rough  surface  of  the  bones,  and  there  appeared  to  have 
been  hyperplasia  of  the  bone  tissue,  the  bones  having  increased  to 
nearly  3  times  the  size  of  the  uninfected  tibiae.  Parallel  punctures 
without  acid  produced  no  such  result. 

All  things  considered,  however,  these  facts  probably  no  longer 
have  significance. 

Rudolph  Wegner  (1876)  reports  that  1:3  or  1:5  lactic  acid 
solution  injected  under  the  periosteum  causes  solution  of  the  bony 
substance,  1:10  acid  showing  no  such  effect. 


554  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Heiss  (1876)  investigated  the  question  as  to  effect  of  lactic 
acid  ingestion  on  the  composition  of  the  bones  of  a  dog.  The  feed- 
ing of  lactic  acid  covered  308  days.  The  body  of  the  dog  was  then 
analyzed.  The  acid  was  administered  at  first  in  quantities  of  1-2 
gm.  daily,  but  during  the  greater  part  of  the  test  in  quantities  of 
7-9  gm.  daily.  The  author  discovered  no  solution  of  the  mineral 
constituents  of  the  bone  by  the  lactic  acid. 

Langendorff  and  Mommsen  (1877)  reported  a  study  of  a  fatal 
case  of  osteomalacia  in  a  man  35  years  old.  Lactic  acid  was  not 
found  in  the  bones. 

Baginsky  (1881)  fed  3  young  dogs,  all  from  the  same  litter,  on 
a  ration  which  was  nearly  free  from  lime.  To  this  diet  was  added, 
with  one  dog,  2  gm.  lactic  acid  per  day,  with  another  2  gm.  calcium 
phosphate,  and  the  third  received  the  basal  ration  alone..  The 
calcium  starvation  caused  by  the  basal  ration  led  to  a  reduction  of 
the  ash  of  the  bones,  and  with  the  dog  receiving  lactic  acid  this 
effect  was  said  to  be  still  further  emphasized. 

Hofmann  (1897)  cites  cases  of  osteomalacia  in  which  no  lactic 
acid  was  present  in  the  urine. 

Klotz  (Jahrb.  f.  Kinderheilk.,  70,  1-61)  found  that  large  doses 
of  lactic  acid  disturb  fat  and  mineral  metabolism.  The  disturb- 
ance of  the  fat  metabolism  may  have  been  due  to  the  neutralization 
of  the  alkali  carbonates  by  the  acid,  with  the  consequent  formation 
of  insoluble  calcium  and  magnesium  soaps.  The  phosphoric  acid, 
no  longer  combined  with  the  calcium  as  insoluble  phosphate,  could 
combine  with  the  alkalis  to  form  soluble  compounds,  thus  increasing 
the  absorption  of  phosphorus. 

Siedamgrotsky  and  Hofmeister  (1879)  published  results  of 
experiments  with  sheep  and  goats  which  received  in  addition  to  a 
ration  of  normal  foods  and  pasture  grass  either  lactic,  sulphuric  or 
hydrochloric  acid.  The  results  are  in  the  nature  of  analyses  of 
the  bones,  balance  experiments  and  analyses  of  the  milk.  In  our 
opinion  the  results  are  inconclusive. 

Moritz  Levy  (1894)  finds  that  lactic  acid  removes  carbonate 
more  rapidly  from  normal  bone  than  it  does  phosphate,  and  that 
therefore  there  is  no  solution  of  bone  salts  by  an  acid  in  osteomal- 
acia.     This  view  of  the  matter  prevails  at  this  time. 

Bonnamour  and  Escallon  (1913)  studied,  with  a  rabbit,  the 
effects  on  the  bones,  of  intravenous  injection  of  lactose.  During  3 
months  one  rabbit  received  378  gm.  of  lactose  in  this  manner  in 
doses  of  10  gm.  each.  Analysis  of  the  bones  revealed  the  follow- 
ing: 


PHOSPHORUS  METABOLISM  555 

COMPOSITION  OF  BONES  AS  AFFECTED  BY  LACTOSE  INJECTION 
Pecent  of  Dry  Bones 

Control  Treated 

CaO  31.66 24.8 

MgO  0.60 3.04 

p205 16.63 25.5 

These  data  imply  a  decalcification  and  an  increase  of  magne- 
sium and  phosphorus. 

COMPOSITION   OP   THE   BONES   IN   OSTEOMALACIA 

A  considerable  number  of  analyses  of  osteomalacic  bones  have 
been  published.  One  of  the  earlier  studies  is  that  of  Mors  and 
Muck  (1869).  Their  figures  show  considerable  variation  in  the 
proportions  of  the  different  mineral  constituents,  with  a  marked 
loss  of  mineral  matter  generally,  and  of  calcium,  a  less  marked  loss 
of  phosphorus,  and  an  increase  in  magnesium.  See  also  von  Gohren 
(1865),  Huppert  (1867)  and  Regnard  (1879). 

The  method  by  which  these  changes  are  produced,  as  stated  by 
Pommer  (1885),  is  through  a  continuous  building  of  new  lime-free 
bone  tissue,  a  cessation  of  the  deposition  of  lime,  and  a  local  removal 
of  lime  from  parts  containing  it.  See  also  von  Recklinghausen 
(1891). 

Moritz  Levy  (1894)  submits  extensive  figures  on  the  composi- 
tion of  osteomalacic  bones,  and  numerous  citations  of  the  work  of 
others.  He  concludes  that  the  relation  6  PO4:10  Ca,  of  normal 
bones,  remains  the  same  during  osteomalacia,  and  that  the  removal 
of  phosphate  takes  place  in  the  same  quantitative  relation  as  that 
of  the  carbonate.  Levy's  own  figures,  however,  show  changes  in 
the  proportion  of  phosphorus  to  calcium  of  various  degrees  up  to 
one-twelfth  of  the  total. 

Galimard  and  Konig  (1905)  publish  analyses  of  bones  from  a 
case  of  infantile  osteomalacia  which  show  a  marked  change  from 
the  normal  relation  of  calcium,  magnesium  and  phosphorus.  The 
following  figures  are  from  this  article. 

PERCENTAGE  COMPOSITION  OF  NORMAL  AND  OSTEOMALACIC 
BONES— Fat-  and  Water-free  Basis 

Normal  Femur  Osteomalacic  Femur 

Ossein 33.35 55.03 

Phosphate  of  lime 55.84 38.07 

Carbonate  of  lime 6.33 6.38 

Phosphate  of  magnesia    0.79 0.49 

McCrudden  (1906a)  found  in  the  osteomalacia  of  horses,  in 
harmony  with  the  conclusions  of  Roloff  (Virchow's  Archiv,  37, 
433),  Huppert  (Archiv  der  Heilkunde,   1866,   1867,   8,   345)    and 


556 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


Chabrie  (Les  phenomene  chimique  de  Fossification,  Paris,  1895,  p< 
65),  that  there  was  a  decrease  of  calcium  and  phosphorus,  and  an 
increase  in  magnesium  and  sulphur.  The  following  data  are  from 
this  article. 

PERCENTAGE  COMPOSITION  OF  RIBS  OF  HORSE,  NORMAL  AND 

OSTEOMALACIC 


Osteomalacic 

Normal 

CaO 

I 

20.09 
0.50 

16.55 
0.35 

II 
18.35 

0.46 
16.00 

0.38 

I 

33.48 
0.11 

23.66 
0.11 

II 
33.12 

MgO 

0.10 

P2O5 

23.22 

s 

0.09 

Cappezzuoli  (1909b)  publishes  analyses  of  long  bones  and  flat 
bones  from  a  case  of  osteomalacia.  He  found  a  general  demineral- 
ization  of  the  bone,  but  a  greater  proportionate  loss  of  calcium  than 
magnesium.  In  the  long  bones  he  found  Ca:Mg:  :100:2.66  and  in 
the  flat  bones  Ca:Mg:  :100:3.85,  while  in  the  normal  the  ratio  of  Ca 
to  Mg  is  as  100:1.14. 

McCrudden  (1910b)  published  a  chemical  analysis  of  bone  from 
a  case  of  human  osteomalacia.      The  results  are  as  follows : 


CaO.. 
MgO.. 

P205.. 
s 


These  figures  show  a  great  decrease  of  calcium,  a  less  marked 
decrease  of  phosphorus,  and  four-fold  increase  of  both  magnesium 
and  sulphur.  While  the  cause  of  the  deposition  of  the  salts  in 
bone  is  not  definitely  known  we  must  admit  (1)  that  there  resides 
in  the  osteogenous  tissue  some  property  which  acts  like  an  affinity 
for  the  bone  salts,  (2)  that  this  affinity  tends  to  maintain  a  constant, 
proportion  between  the  salts  deposited,  but  (3)  that  variations  in 
the  composition  and  reaction  of  the  blood,  as  affected  by  disease, 
modify  the  character  of  this  affinity,  as  indicated  by  the  composi- 
tion of  the  bones,  in  definite  and  consistent  ways,  even  if  not  to 
unlimited  degrees. 

METABOLISM  IN  OSTEOMALACIA 

Mohr,  in  von  Noorden's  "Metabolism  and  Practical  Medicine," 
tabulates  calcium,  magnesium  and  phosphorus  balances  by  von  Lim- 
beck (1894),  S.  Neumann  (1894a,  1894b)  von  Korczynski  (1902), 


PHOSPHORUS  METABOLISM 


557 


Sauerbruch  (1902),  Goldthwait  et  al.  (1905),  and  Hotz  (1906). 
Improved  physical  condition  of  the  patients  was  usually  accom- 
panied by  increased  retention  of  the  bone  salts.  Phosphorus  reten- 
tion was  commonly  disproportionately  large,  as  compared  with 
calcium,  and  both  may  be  retained  in  moderately  severe  cases. 

Von  Korczynski  (1902)  reports  balance  data  with  two  cases  of 
osteomalacia  snowing  that  if  the  disease  is  not  too  far  advanced, 
and  the  course  of  the  disease  not  rapid,  and  no  marked  cachexy  is 
apparent,  the  organism  retains  phosphoric  acid. 

Earlier  metabolism  studies  showed  considerable  losses  of  cal- 
cium, magnesium  and  phosphorus,  not  only  in  the  urine  but  also  in 
the  feces,  in  bronchial  mucous  in  cases  associated  with  bronchial 
catarrh,  and  also  in  the  milk.  See  Pagenstecher  (1862),  Gusserow 
(1862),  Schmuziger  (1875)  and  von  Limbeck  (1894). 

Raspopoff  (1884)  finds  the  proportion  of  the  urinary  phospho- 
rus united  to  alkaline  earths  greater,  and  that  united  to  the  fixed 
alkalis  less,  in  osteomalacia  than  the  normal.  See  also  Raspopoff 
(1885). 

Metabolism  studies  by  S.  Neumann  (1894a,  1894b)  show  that  a 
predominating  loss  of  phosphorus  may  change  to  as  considerable  a 
retention  of  the  same  as  the  case  improves,  while  on  the  other  hand 
the  work  of  His  (1902),  on  a  case  of  infantile  osteomalacia,  shows 
that  improvement  may  be  due  to  improved  retention  of  calcium,  the 
preexisting  positive  phosphorus  balance  remaining  without  striking 
change. 

The  existence  of  typical  osteomalacia  in  children  was  attested 
by  investigations  of  Siegert  (1898),  who  cites  a  previous  demon- 
stration by  von  Recklinghausen,  and  also  by  the  above-mentioned 
work  of  His. 

Very  few  studies  of  the  blood  in  osteomalacia  have  come  to  our 
attention.  Fehling  (1891)  reports  subnormal  alkalinity  in  the 
blood  in  severe  cases;  Pende  (1905)  found  improved  blood  condi- 
tions as  determined  by  corpuscle  count,  etc.,  after  phosphorus 
therapy.  Kobler  (1888)  also  publishes  blood  ash  analyses  contrasted 
with  the  normal,  with  results  as  follows : 


COMPOSITION  OF  ASH  OF  BLOOD 

Osteomalacia 

Normal  (Jarisch, 
mean  of  four) 

7.2 

16.04 
19.925 
34.160 
9.35 
0.52 

12.85 

8.49 

Sulphuric  anhydride 

6.85 

29.59 

25.565 

Soda 

23  169 

0.872 

0.512 

7.86 

558  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

A  somewhat  recent  idea  as  to  the  cause  of  osteomalacia  is  that 
of  Hoennicke  (1904,  1905)  (10  pp.  bibliog.),  who  considers  it  due  to 
disease  of  the  thyroid  gland. 

Zunz  (1913)  found  in  osteomalacia  no  characteristic  variation 
in  protein  metabolism.  The  phosphorus  metabolism  was  quite 
variable,  but  a  high  fecal  outgo  was  rather  constant,  this  elimina- 
tion being  usually  reduced  by  ovariotomy,  with  increased  tendency 
to  retention. 

TBEATMENT  OF  OSTEOMALACIA 

In  the  treatment  of  osteomalacia  His  (1902)  notes  favorable 
results  from  the  use  of  phosphorus,  first  by  Trousseau  in  1868, 
later  by  Wegner  in  1872,  cites  Sternberg's  report  (1893)  of  eight 
cured  cases  from  the  clinics  of  Busch,  Strumpell,  Matterstock  and 
Nothnagel,  and  also  his  own  description  of  four  other  such  cases, 
also  favorable  results  by  von  Limbeck  (1894),  Kosminski  (1896), 
Bernstein  (1898),  Heubner  (1898),  Siegert  (1898)  and  Littauer 
(1899).  He  also  cites  unfavorable  reports  of  the  use  of  phospho- 
rus in  osteomalacia  by  Gelpke  (1891),  and  Fehling  and  Wetzel 
(1899). 

Other  favorable  reports  from  the  use  of  phosphorus  in  osteo- 
malacia have  been  published  by  Warschauer  (1890),  Sternberg 
(1893),  H.  Fischer  (1894),  Pende  (1905)  and  Hotz  (1906). 

Treatment  with  oophorin  and  thyraden  by  Senator  (1897)  led 
to  improvement  in  the  patient,  but  treatment  with  oophorin  by 
Bernstein  (1898)  failed,  after  which  phosphorus  therapy  caused 
a  cure. 

Ovariotomy  has  given  good  results  in  a  number  of  cases  but 
there  is  no  established  connection  between  the  ovaries  and  osteo- 
malacia. Beneficial  results  seem  to  be  due  especially  to  the  preven- 
tion of  loss  of  calcium  and  phosphorus  from  the  body  through  the 
various  incidents  of  the  sexual  life. 

Fehling  (1891)  reports  cases  of  ovariotomy,  and  later  (1895) 
10  cures  from  12  such  operations.  Curatulo  and  Tarulli  (1895) 
also  report  favorable  results  from  ovariotomy. 

S.  Neumann  (1896)  reported  metabolism  data  with  cases  of 
ovariotomy  in  osteomalacia.  This  treatment  is  regarded  as  radical, 
and  not  generally  to  be  recommended,  though  benefit  resulted  in 
case  the  disease  was  not  too  severe.  Results  were  unfavorable  in 
advanced  cases. 

Goldthwait,  Painter,  Osgood  and  McCrudden  (1905)  made  bal- 
ance experiments  on  a  16-year-old  osteomalacic,  both  before  and 
after  ovariotomy.      Before  the  operation  there  was  a  marked  loss 


PHOSPHORUS  METABOLISM  559 

of  calcium,  a  slight  loss  of  phosphorus,  and  a  slight  storage  of  mag- 
nesium. Some  months  after  the  operation  the  patient  was  found 
to  be  storing  calcium,  but  magnesium  and  phosphorus  determina- 
tions were  not  made.  These  authors  consider  that  their  balances 
indicate  a  partial  replacement  of  calcium  by  magnesium  previous  to 
the  operation. 

McCrudden  (1906b)  reports  a  later  metabolism  study  on  the 
same  case  of  osteomalacia  that  was  studied  a  year  and  a  half  before 
by  Goldthwait  et  al,  and  found  much  improved  by  ovariotomy.  At 
the  time  of  this  later  study  the  patient  was  once  more  in  a  critical 
condition,  and  was  found  to  be  losing  calcium  and  magnesium,  but 
retaining  phosphorus.  As  to  the  connection  of  the  ovaries  with 
osteomalacia  it  was  McCrudden's  idea  that  ovariotomy  is  without 
influence  on  the  ultimate  cause  of  the  disease. 

In  accord  with  this  idea  we  have  the  results  of  Luthje  (1903), 
who  showed  by  the  removal  of  testes  and  ovaries  from  dogs  that 
neither  organs  bear  any  noticeable  relation  to  phosphorus  metabo- 
lism generally. 

Curshmann  (1911)  describes  cases  of  osteomalacia  in  men  and 
women  of  middle  age  and  old  age.  Cure  was  effected  by  the  use  of 
phosphorized  cod-liver  oil. 

In  this  connection  we  have  also  considered  the  paper  of  Ogata 
(1911-12). 

PHOSPHORUS  METABOLISM  AND  THE  PARATHYROIDS 

The  extirpation  of  the  parathyroids  reveals  an  essential  relation 
of  these  glands  to  metabolism,  the  nature  of  which  is  as  yet  but 
imperfectly  understood.  The  following  notes  show  that  they  are 
intimately  connected  with  phosphorus  metabolism. 

Morel  (1910)  notes  that  parathyroidectomized  animals  fail  to 
heal  fractures  as  rapidly  as  normal  animals,  the  conversion  of  carti- 
lage to  bone  being  slower.  Removal  of  the  thyroid  was  without 
appreciable  effect  on  the  bones.  Also  (1909)  administration  of 
thyroid  substance  was  without  effect  on  the  bones  of  adult  animals, 
while  the  feeding  of  parathyroid  extract  to  young  rabbits  favored 
the  growth  of  the  bones  independently  of  the  calcium  of  the  ration. 

Erdheim  (1911)  found  that  in  rats,  after  parathyroidectomy, 
the  new  bone  produced  is  very  poor  in  calcium,  and  that  the  skeleton 
generally  takes  on  an  osteomalacic  or  rachitic  character. 

T.  Bauer  (1911)  supports  Erdheim's  findings  of  parathyroid 
hyperplasia  in  human  osteomalacia,  which  hyperplasia  is  ascribed, 
to  an  increased  functional  demand  for  calcium. 


560 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


Erdheim  also  finds  that  after  parathyroidectomy  the  dentine 
calcifies  late,  or  not  at  all;  that  the  enamel  is  deficient,  and  that 
fractures  of  the  bones  are  frequent,  but  that  if  parathyroids  be 
successfully  transplanted  into  these  parathyroidectomized  animals 
the  dentine  calcifies;  if  they  are  then  removed  the  dentine  forms 
without  calcium. 

Toyofuku  (1911)  made  a  histological  study  showing  that 
deficient  deposition  of  calcium  is  responsible  for  the  structural 
change  observed. 

These  above  observations  regarding  calcium  involve,  of  course, 
phosphorus,  without  which  calcium  could  not  be  used  in  the  normal 
growth  of  bone. 

Greenwald  (1911,  1913a)  found  that  parathyroidectomy 
caused  a  very  marked  phosphorus  retention,  which  was  followed, 
but  not  until  the  appearance  of  tetany,  by  an  increased  excretion  of 
phosphorus.  In  another  paper  (1913b)  Greenwald  reported  that 
after  parathyroidectomy  the  phosphorus  of  the  blood  is  increased, 
the  greater  part  of  this  increase  being  in  the  fraction  which  is 
insoluble  in  the  usual  lipoid  solvents,  but  is  soluble  in  a  mixture  of 
dilute  hydrochloric  or  acetic  and  picric  acids.  Some  of  Greenwald's 
data  are  as  follows : 

PHOSPHORUS  IN  BLOOD  FROM  A  THYRO-PARATHYROIDECTOMIZED 
DOG  AND  A  NORMAL  DOG— Milligrams  of  Phosphorus  per  Kilo 

of  Blood 


Phosphorus 

Parathy- 
roidectomized 
214 

Normal 
214B 

Total 

436 

162 

8 

233 

33 

370 

140 

Extracted,  with  alcohol  and  ether 

7 

192 

31 

PHOSPHORUS  IN  SERUM  FROM  PARATHYROIDECTOMIZED  AND 
NORMAL  DOGS— Milligrams  of  Phosphorus  per  Kilo  of  Serum 


Phosphorus 

Parathyroidectomized 

Normal 

Dog-  215 

Dog-  217 

Dog-  219* 

Dog  216 

Dog  218 

Dog  220 

Total 

In  acetone  extract. 
In  acid  extract 

212 
149 
62.3 

222 
128 
76.7 

291 
194 
87.7 
3.8 

157 
131 
26.9 

244 

186 
44.4 
2.4 

167 
110 
45.5 

*   Complete  thyroidectomy  in  this   case. 


PHOSPHORUS  METABOLISM  561 

Paladino  (1913)  considers  that  after  the  removal  of  the  thyroid 
and  parathyroid  glands  urinary  phosphorus  elimination  increases 
greatly,  but  Greenwald  (1913c)  states  that  the  removal  of  these 
organs  reduces  phosphorus  elimination,  and  that  there  is  no  increase 
until  the  appearance  of  tetany,  the  increase  then  being  due  to  the 
muscular  activity. 

PHOSPHATURIA 

Phosphaturia  is  not  a  disease;  it  is  a  symptom  or  condition 
which  may  result  from  many  causes,  normal  as  well  as  pathological. 
It  is  said  to  exist  when,  from  whatever  cause,  there  is  a  sediment 
of  alkaline  earth  phosphates,  which  may  be  mixed  with  carbonates, 
in  the  fresh  urine.  The  immediate  cause  of  the  sediment  is  a 
relative  increase  of  alkalis  or  alkaline  earths,  or  a  relative  decrease 
of  acid  elements.  In  either  case  the  increase  or  decrease  may  be. 
relative  without  being  absolute;  hence  the  existence  of  the  phos- 
phate sediment  is  no  indication  of  the  amount  of  phosphorus  in  the 
urine.  It  may  occur  with  increased  or  decreased  phosphorus  so 
long  as  there  is  sufficient  relative  increase  of  alkali  or  alkaline 
earth. 

The  method  of  formation  of  the  sediment  is  a  change,  under 
the  influence  of  alkalis  or  alkaline  earths,  of  a  portion  of  the  soluble 
diacid-phosphates  normally  present  into  the  insoluble  monacid  or 
normal  phosphates. 

This  condition  may  result  from  any  such  change  of  diet  as 
increases  to  a  sufficient  extent  the  carbonates  or  oxidizable  organic 
compounds  containing  alkalis  or  alkaline  earths ;  or  through,  any 
such  influence  as  decreases  the  acids  normally  excreted  in  the  urine, 
such  circumstances  for  instance  as  the  ingestion  of  much  protein, 
or  continued  washing  out  of  the  stomach,  both  of  which  result  in  the 
separation  from  the  blood  of  unusual  amounts  of  hydrochloric  acid, 
and  liberate  within  the  blood  corresponding  amounts  of  alkali. 

Another  form  of  phosphaturia,  common  in  children,  is  due  to 
a  deflection  of  calcium  from  feces  to  urine  without  necessary  change 
in  the  amount  of  either  calcium  or  phosphorus,  the  cause  not  being 
established  beyond  possibility  of  doubt,  but  commonly  thought  to  be 
such  pathological  condition  of  the  large  intestine  as  results  in 
decreased  ability  to  excrete  calcium. 

Aside  from  matters  of  diet  and  digestive  disorder  it  seems 
likely  that  nervous  influences  affect  metabolism  in  such  ways  as  to 
cause  phosphaturia,  either  combined  or  not  with  the  juvenile  form 
above  described;  and  phosphaturia  is  common  in  many  diseases; 
for  instance,  in  some  cases  of  diabetes  mellitus,  in  tuberculosis,  and 
in  such  bone  diseases  as  ostitis  and  osteomvelitis. 


562  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

It  is  impossible  for  us  to  attempt  a  review  of  the  enormous 
literature  of  this  subject,  but  we  shall  record  observations  on  a  few 
of  the  more  important  articles  which  have  come  to  our  attention. 

One  of  the  more  important  of  the  earlier  works  on  phosphaturia 
is  that  of  Teissier  (1875,  1877).  He  found  that  it  was  a  condition 
not  particularly  connected  with  alterations  in  skeleton  or  nerves. 
A  part  of  his  conclusions  are  as  follows : 

"Permanent  phosphaturia  always  indicates  serious  trouble  in 
the  general  nutrition.  The  trouble  may  be  profound  enough  to 
show  all  the  apparent  symptoms  of  sugar  diabetes.  But  phos- 
phaturia of  the  diabetic  form  is  not  an  essential  disease,  but  js 
rather  a  morbid  complication  which  may  be  sympathetic  of  various 
affections.  It  may  be  a  simple  form  of  nervousness  which  accom- 
panies pulmonary  phthisis,  or  may  be  a  precursory  sign  of  it.  It 
may  be  symptomatic  of  sugar  diabetes,  latent  or  transformed. 
Diabetic  phosphaturia  may,  like  sugar  diabetes,  exert  an  evil  influ- 
ence on  the  progress  of  traumatic  lesions." 

"Phosphates  are  abundant  in  the  urine  of  consumptives  at  the 
beginning  of  the  disease ;  they  diminish  somewhat  as  the  period  of 
tuberculous  cachexy  arrives.  They  diminish  in  true  chlorosis. 
They  increase  in  diseases  of  the  brain,  and  of  the  marrow.  They 
increase  in  chronic  rheumatism.  They  decrease  generally  in  the 
course  of  fever  and  ague.  They  do  not  increase  in  spite  of  most 
abundant  feeding  in  the  course  of  convalescence;  rather  they  are 
diminished." 

Ralfe  (1887)  classified  cases  of  phosphaturia  coming  to  his 
attention  (1)  as  associated  with  disturbances  of  the  nervous  system, 
(2)  as  associated  with  pulmonary  disease,  (3)  as  alternating  or 
coexisting  with  saccharine  diabetes  and  (4),  as  without  special  con- 
nection. 

Sendtner  (1888)  showed  in  a  single  case  that  excess  of  lime 
was  the  cause  of  phosphaturia. 

Peyer  (1889)  also  considered  excess  alkalinity  of  the  blood  as 
the  cause  of  phosphaturia. 

Klemperer  (1899)  considered  phosphaturia  as  due  to  hyper- 
acidity and  motor  insufficiency  of  the  stomach,  the  former  taking 
abnormal  amounts  of  acid  from  the  blood,  and  the  latter  by  retain- 
ing it  over-long  in  the  stomach,  delaying  its  return  in  compensating 
amount  by  way  of  intestinal  absorption.  Klemperer  considered 
both  of  these  symptoms  as  due  to  nervous  affection. 


PHOSPHORUS  METABOLISM  563 

Leo  (1902)  reported  a  case  of  phosphaturia  in  which  there  was 
abundant  phosphate  and  carbonate  precipitate  in  the  urine,  the 
same  apparently  not  being  due  either  to  motor  insufficiency  or 
hyperacidity  of  the  stomach,  since  the  stomach  secretion  was  alkal- 
ine at  night  and  early  morning. 

Soetbeer  (1902)  made  a  very  satisfactory  comparison  of  a  child 
exhibiting  phosphaturia  with  one  in  normal  condition.  The  phospho- 
rus in  the  urine  was  about  the  same,  slightly  higher  with  the  healthy 
child,  but  the  lime  more  than  two-and-a-half  times  as  much  in  the 
urine  of  the  patient  as  in  that  of  the  control.  The  phosphorus  of 
the  feces  was  the  same  in  both  cases,  as  also  was  the  lime  insoluble 
in  water  but  soluble  in  hydrochloric  acid,  while  the  water-soluble 
lime  was  0.037  gm.  in  the  patient  and  0.310  gm.  in  the  control;  thus 
the  essential  difference  in  these  cases  was  that  the  control  had 
excreted  water-soluble  lime  into  the  intestine,  while  the  patient  had 
excreted  the  same  into  the  urine,  the  difference  apparently  being 
due  to  intestinal  catarrh  in  the  pathological  case. 

Soetbeer  and  Krieger  (1902)  reported  a  case  of  phosphaturia 
where  the  ratio  P205:CaO  was  sometimes  1.5  or  2  to  1  instead  of 
12  to  1,  the  normal  relation,  while  the  urinary  elimination  of  lime 
rose  to  almost  0.7  gm.  daily,  the  normal  being  about  0.2  gm.  See 
also  Panek  (1900)  and  De  Lange  (1903). 

Gouraud  (1903)  investigated  the  ratio  of  P  to  N-  in  the  urine  of 
patients  suffering  from  pneumonia,  typhoid  fever,  rheumatism, 
cerebrospinal  meningitis,  tubercular  meningitis,  diabetes,  neuras- 
thenia and  false  and  true  phosphaturia.  No  account  was  taken 
of  food  or  feces  phosphorus.  Gouraud  distinguishes  between 
false  phosphaturia,  ordinarily  due  to  alkalinity  of  the  urine,  and  true 
phosphaturia,  due  especially  to  (1)  dyspepsia,  especial- 
ly hyperchlorhydria,  which  produces  only  a  minimum  increase, 
(2)  tuberculosis,  with  phosphaturia  as  a  complication,  (3) 
diabetes,  in  its  more  serious  forms,  and  (4)  nervous  states,  partic- 
ularly neurasthenia. 

Von  During  (1905)  studied  60  cases  of  phosphaturia.  He 
concluded  that  cases  of  phosphaturia,  attributed  to  neurasthenia, 
were  actually  caused  by  urethritis  and  prostatitis,  the  neurasthenia 
being  secondary,  and  sometimes  late  in  appearing.  Von  During 
explains  it  by  the  idea  that  the  whole  urogenital  tract  is  inflamed  by 
"inflammation  of  any  part,  the  parenchyma  of  the  kidneys  thus 
becoming  impaired.  Albuminuria  was  found  often  to  accompany 
phosphaturia. 


664  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Moll  (1905)  describes  a  case  of  hysteria  combined  with  intes- 
tinal catarrh  in  a  child  51/2  years  old.  A  symptom  of  the  case  was 
an  abnormal  amount  of  calcium  phosphate  in  the  urine.  While  in 
this  condition  the  child  lost  2.5  kg.  in  weight  in  2.5  months.  After 
a  change  of  diet,  and  an  avoidance  of  lime-rich  foods  there  was  an 
improvement  in  general  health,  and  a  gain  in  weight  of  2  kg.  in  V/% 
months,  the  urine  being  normal.  With  a  return  to  the  original 
diet  came  a  return  of  the  old  symptoms,  and  an  increase  of  calcium 
and  phosphorus,  especially  the  former,  in  the  urine.  The  diet  was 
once  more  changed  to  fat,  meat,  sugar  and  fruits,  thus  avoiding 
lime,  with  the  result  that  the  urine  became  normal,  and  the  nervous 
and  digestive  disturbances  disappeared. 

Moll  (1909)  published  an  extensive  report  of  further  studies  of 
this  same  subject,  having  reference  to  breast-fed  infants.  In  this 
study  he  made  a  comparison  of  the  composition  of  the  urine  of 
healthy  infants  and  those  suffering  from  defective  digestion,  the 
following  are  a  part  of  the  conclusions  from  parts  I  and  II  of  the 
report : 

The  urine  of  healthy  breast-fed  infants  contains  no  phosphorus, 
or  only  traces. 

The  dyspeptic  infant  excretes  more  or  less  phosphorus  through 
the  urine.  This  amount  may  be  decreased  by  a  short  fasting  period 
(24  hours),  a  reduction  of  the  number  of  feedings,  or  artificial 
evacuation  of  the  intestine.  If  the  trouble  is  relieved  the  phospho- 
rus content  of  the  urine  decreases;  if  it  continues  the  phosphorus 
increases. 

In  inanition,  in  the  ordinary  sense  of  the  word,  if  not  carried 
too  far,  no  phosphorus,  or  only  a  trace  is  excreted  in  the  urine  of 
the  healthy  child. 

If  a  breast-fed  child  gains  in  weight  and  shows  no  sign  of  ill- 
ness, and  has  no  phosphorus  in  the  urine,  then  it  is  healthy. 

If  the  breast-fed  infant  gains  in  weight,  and  with  insufficient 
food  intake  shows  no  phosphorus,  or  only  traces,  in  the  urine,  it 
is  starved. 

If  the  child  does  not  gain  or  lose  in  weight,  and,  with  appar- 
ently insufficient  nourishment,  excretes  large  quantities  of  phospho- 
rus in  the  urine,  it  has  a  digestive  disorder.  Thus  the  content  of 
the  urine  in  phosphorus  is  a  gauge  of  the  condition  of  the  infant. 

Loss  in  weight  and  a  high  percentage  of  phosphorus  in  the 
urine  run  in  a  parallel  way ;  also  gain  in  weight  and  a  low  percent- 
age of  phosphorus  in  the  urine. 


PHOSPHORUS  METABOLISM  565 

The  same  milk  reacts  differently  on  different  children.  It  is 
possible  then  to  determine  the  effect  on  the  child  of  a  change  of  milk 
by  testing  the  urine  (24  hours)  for  phosphates.  If  benefit  has 
been  received  the  urine  will  show  a  considerable  decrease  in  phos- 
phorus. 

In  part  III  of  this  article  Moll  discusses  acute  gastroenteritis 
in  infants,  and  submits  experimental  data  showing  the  effect  of 
treatment  on  the  phosphorus  of  the  urine. 

In  the  height  of  the  disease  there  is  a  high  percentage  of  phos- 
phorus in  the  urine,  which  is  quickly  decreased  by  stopping  the  food 
intake,  and  substituting  a  water  diet.  These  low  values  persist 
during  convalescence.  When  breast-feeding  is  resumed  the  values 
of  urine  phosphorus  increase  again.  The  tolerance  of  the  organ- 
ism toward  new  food  is  measured  by  the  urine  analysis.  The  organ- 
ic phosphorus  present  in  severe  stages  of  the  disease  disappears 
with  the  decrease  of  total  phosphorus  in  the  urine. 

In  cases  which  have  a  fatal  termination  this  fasting  therapy 
does  not  produce  the  above  results,  that  is,  improvement,  in  general, 
accompanied  by  decreased  urine  phosphate. 

Organic  phosphorus  in  the  urine  of  the  breast-fed  child  is  to  be 
regarded  as  a  pathological  symptom. 

Von  Moraczewski  (1905)  gave  attention  to  the  balance  of  acid 
and  basic  elements  in  pathological  urines.  His  table  as  published 
is  in  need  of  recalculation.  A  part  of  the  conclusions  are  as  follows : 

The  chronic  phosphaturia  which  does  not  show  the  clinical 
symptoms  of  neurasthenic  phosphaturia,  or  of  the  phosphaturia  of 
children,  is  characterized  by  an  abnormal  relation  of  phosphorus 
and  calcium  elimination  in  the  urine,  in  that,  coincident  with  a 
decrease  of  lime  salts  there  is  an  increase,  though  small,  of  phos- 
phorus. 

By  treatment  with  alkalis  the  elimination  of  the  metalloid  ions 
(chlorine,  sulphur  and  phosphorus)  is  relatively  more  favored  than 
that  of  the  metal  ions,  by  which  the  relative  elimination  returns  to 
the  normal.  One  and  the  same  individual  may  have  this  form  of 
phosphaturia  and  gout ;  that  is,  (a)  may  show  phosphaturia  alter- 
nating with  oxaluria  or  gout;  (b)  may  suffer  generally  with  phos- 
phaturia and  then  with  gout,  or  the  reverse.  Normal  urine  contains 
more  acid  ions  than  the  urine  of  phosphaturia  or  gout. 

Tobler  (1905)  published  data  on  two  cases  of  phosphaturia. 
As  usual  the  lime  in  the  urine  was  high,  while  the  phosphorus 
remained  within  normal  limits,  but  the  lack  of  food  analyses  limits 
the  use  of  the  data. 


566  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Doctor  (1906)  says  that  phosphorus  estimations  show  that  it  is 
incorrect  to  designate  every  case  of  alkalinuria  as  phosphaturia. 

Langstein  (1906)  observes  the  presence  of  phosphaturia  in 
cases  of  disease  where  there  was  not  the  usual  excess  alkali 
or  excess  removal  of  acid  as  a  cause.  He  mentions  sexual  phos- 
phaturia, due  to  a  complication  of  diseases  of  the  urogenital  system ; 
neurasthenic,  causing  reflex  effects  on  kidney  secretions ;  that  form 
which  von  Moraczewski  describes  as  associated  with  uric  acid 
diathesis,  and  finally  the  juvenile  form.  Langstein  describes  in 
detail  the  nervous  symptoms  accompanying  the  juvenile  form.  He 
considers  overnourishment  on  foods  such  as  milk  and  eggs,  which 
are  rich  in  lime  to  be  the  cause,  and  found  that  dietetic  changes  on 
this  basis  caused  marked  improvement.  In  his  cases  there  was  no 
intestinal  catarrh  demonstrable.  Langstein  emphasizes  the  diver- 
sity of  causes  of  phosphaturia. 

Novi  (1908)  reports  a  reduction  of  the  urinary  phosphorus,  in 
phosphaturia  following  antirabes  treatment,  from  69.11  percent  of 
an  intake  of  2.457  gm.  to  46.07  percent  of  an  intake  of  2.451  gm., 
plus  0.603  gm.  P205  in  the  shape  of  phytin,  the  difference  being 
ascribed  to  this  phosphatic  supplement. 

Klemperer  (1908),  in  an  extended  study  of  various  phases  of 
phosphaturia,  found  that  doses  of  0.3  gm.  oxalic  acid  plus  3.0  gm. 
NaHCOg  greatly  decreased  the  absorptive  power  of  the  kidneys  for 
calcium,  as  also  did  very  small  quantities  of  HgCl2  (1  drop  3  times 
daily  of  a  5-percent  solution,  in  all  1.0  mg.  in  24  hours). 

PHYTIN  THERAPY 

This  subject  is  discussed  in  connection  with  normal  phytin 
metabolism;  see  p.  315. 

RACHITIS 

Rachitis  has  been  known  for  a  very  long  time;  at  least  it  was 
described  by  Glissonius  in  1650.  It  is  a  disease  of  infants  which 
involves  phosphorus  metabolism,  especially  through  its  effects  upon 
the  bones,  but  also  apparently  in  other  ways  as  well.  In  the  literature 
of  the  subject  much  prominence  is  given  to  the  element  calcium, 
but  we  may  bear  in  mind  the  fact  that  one  could  as  consistently  put 
the  same  emphasis  on  phosphorus,  for  these  two  elements  are  used 
together  in  the  tissues  most  involved  in  rachitis. 

In  rickets  calcification  of  the  bones  is  irregular;  areas  of  partial 
ossification  lying  between  areas  of  greatly  enlarged  cartilage  cells. 
The  bone  is  deeply  imperforate  by  projections  into  it  of  both  mar- 
row and  periosteum,  and  the  marrow  spaces  are  enlarged  and  irreg- 
ular. Absorption  of  formed  bone  is  not  prominent  as  in  osteomal- 
acia, but,  rather,  a  failure  of  the  osteoid  tissues  to  calcify. 


PHOSPHORUS  METABOLISM  567 

Associated  with  rickets  there  are  often  gastrointestinal  disor- 
ders, and  anaemic  changes  in  the  blood.  The  blood  alkalinity,  how- 
ever, was  found  normal  by  Stoltzner. 

The  largest  part  of  the  literature  of  rachitis  has  to  do  with  its 
etiology.  This  aspect  of  the  problem,  however,  remains  unsolved. 
Among  the  possible  causes,  lack  of  calcium  in  the  food,  deficient 
absorption  of  calcium,  and  acid  intoxication,  none  fits  the  facts  in  a 
thoroughly  satisfactory  manner.  From  an  enormous  literature,  of 
which  we  do  not  attempt  to  review  the  whole,  the  following  are 
some  representative  fragments. 

Among  the  earlier  papers  which  we  have  seen  are  those  of 
Gobley  (1844, 1846),  who  used  ray's  liver  oil  with  success  in  rickets, 
Virchow  (1853),  who  histologically  distinguished  rickets  from 
osteomalacia,  G.  Wegner  (1872),  who  studied  the  effects  of  the 
element  phosphorus  repeatedly  administered  in  small  doses,  and 
suggested  the  probable  benefit  from  such  treatment  in  rachitis, 
Heitzmann  (1873),  who  thought  he  produced  rickets  and  osteomala- 
cia with  lime-poor  food  and  lactic  acid,  and  Roloff  (1875a,  1875b, 
1879),  who  also  produced  and  cured  artificial  rickets,  and  considered 
it  actual  rickets,  which  he  thought  due  to  lack  of  phosphorus  in  the 
food. 

Seeman  (1879)  considered  rickets  as  caused  by  deficient  absorp- 
tion of  lime,  due  to  digestive  disorders  originating  in  a  subnormal 
hydrochloric  acid  content  of  the  gastric  juice. 

Zander  (1881)  also  had  the  idea  of  a  deficiency  of  hydrochloric 
acid  in  the  gastric  juice,  and  considered  this  due  to  an  excess  of 
potassium  and  phosphorus  in  the  mother's  milk  causing  an  undue 
elimination  of  sodium  and  chlorine.  These  observations  were 
based  on  the  study  of  the  composition  of  the  milk  of  the  mothers  of 
normal  and  of  rachitic  children. 

E.  Voit  (1880)  recognized  the  futility  of  feeding  calcium  salts 
in  actual  rickets,  and  recommended  attention  to  the  gastrointestinal 
disorders  often  accompanying  rickets,  considering  these  the  cause, 
and  their  relief  the  cure. 

Kassowitz  (1881,  1884a,  1884b,  1886)  experimented  with  phos- 
phorus therapy  in  rickets.  He  considered  that  it  reduced  the 
morbidly  increased  vascularization  of  the  osteoid  tissue  from  which 
result  the  characteristic  bone  conditions.  Kassowitz  experimented 
with  many  thousands  of  cases  (1890)  and  recommended  "lipanin," 
olive  oil  and  phosphorus.  Other  statements  of  his  ideas  and  results 
are  found  in  his  publications  of  1901, 1910, 1912.  He  expresses  the 
idea  that  rachitis  is  not  due  to  lack  of  lime  in  the  food.  Phosphorus, 


568  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

he  finds,  causes  hardening  of  the  bone  through  the  laying  on  of  lime 
salts,  and  he  speaks  of  rickets  as  involving  an  abnormal  increase  of 
tissue  building,  and  an  abnormal  fullness  of  blood  in  the  bone- 
making  tissues. 

J.  Cohnheim  (1882)  published  an  extended  discussion  of  the 
disease,  as  also  did  Pommer  (1885),  who  considered  it  as  originating 
outside  the  skeleton. 

Baginsky  (1882)  studied  demineralization  by  lactic  acid  in  con- 
nection with  rickets. 

Anna  Schabanowa  (1889)  reported  successful  use  of  phospho- 
rus in  rickets. 

Korsakov  (1892)  could  not  cause  rickety  conditions  by  feeding 
lactic  acid.  Phosphorus  therapy  caused  varying  results,  some 
successful,  others  not.  Strontium  salts  used  in  rickets  caused  a 
certain  amount  of  ossification.  Cremer  (1891),  however,  found 
that  strontium  phosphate  would  not  prevent  the  appearance  of 
rickety  symptoms  in  a  dog  on  a  low  calcium  diet,  though  it  was 
deposited  to  some  extent  in  the  bones. 

Brubacher  (1890)  shows  that  in  congenital  rickets  the  body  is 
characteristically  poor  in  ash.  On  the  dry,  fat-free  basis  calcium 
and  phosphorus  are  both  markedly  deficient,  the  former  decidedly 
more  so  than  the  latter.  The  analysis  of  the  ash  shows  calcium 
quite  deficient,  and  phosphorus  only  slightly  so.  Magnesium  seems 
not  to  vary  in  an  important  way.  Brubacher  agreed  with  Kassowitz 
in  the  belief  that  organs  other  than  the  bones  were  not  subnormal 
in  calcium. 

Herter  (1898)  experimented  with  fat-free  rations  on  swine, 
having  in  mind  the  alleged  low  fat  content  of  the  milk  of  women 
whose  children  have  grown  rachitic  on  the  mother's  milk,  and  also 
the  occurrence  of  rickets  in  children  fed  largely  on  condensed  milk. 
He  concluded  that  fat  starvation  causes  a  very  imperfect  absorption 
of  phosphorus  from  the  intestine;  but  since  complete  balance  data 
were  lacking  we  must  suspend  judgment  on  this  matter.  There 
was  nothing  resembling  or  suggesting  rickets,  however,  in  the 
results  from  the  fat-free  rations. 

Jviidel  (1893b)  satisfactorily  demonstrated  that  lime  was 
absorbed  from  the  carbonate  and  the  acetate,  and  was  also  excreted, 
by  rickety  children,  in  a  normal  manner. 

Chabrie  (1894,  1895)  developed  an  interesting  theory  of  the 
causation  of  rickets  by  the  interference  of  lactic  acid  in  the  normal 
formation  of  collagen  from  chondrin,  a  process  necessary  to  normal 
ossification,  and  also  the  interference  of  lactic  acid  with  the  normal 
precipitation  of  calcium  phosphate  and  calcium  carbonate  in  bone  by 


PHOSPHORUS  METABOLISM 


569 


the  fixation  by  lecithin  of  the  carbonic  acid  holding  them  in  solution, 
lecithin  thus  being  necessary  to  normal  utilization  of  inorganic 
phosphates.  For  details  see  the  original.  He  also  explained  the 
partial  replacement  of  calcium  of  the  bones  by  magnesium,  in  osteo- 
malacia, by  reference  to  a  different  relation  to  lecithin. 

Flieger  (1897)  reported  many  unsuccessful  attempts  to  cure 
rickets  with  phosphorus  in  cod-liver  oil.  The  reason  for  his  fail- 
ure is  not  apparent. 

Zweifel  (1900)  advanced  the  theory  that  deficient  hydrochloric 
acid  of  the  gastric  juice,  as  caused  by  lack  of  common  salt  in  the 
diet,  interfered  with  the  solution  of  calcium  salts,  and  so  caused 
rickets;  and  Delcourt  (1899)  stated  that  he  had  produced  in  bones 
lesions  similar  to  those  of  rickets  by  means  of  potassium  phosphate. 

Leichnam  (1903)  found  phosphorus,  calcium,  magnesium  and 
chlorides  all  above  normal,  and  urea  below  normal,  in  the  urine  in 
rickets,  indicating  a  profound  state  of  denutrition. 

W.  Stoeltzner  (1903)  reports  negative  results  from  phosphorus 
therapy  in  much  complicated  cases.  He  recommended  phosphorus, 
however,  (1904)  to  be  administered  in  cod-liver  oil,  and  suggested 
as  a  cause  of  rickets  the  functional  insufficiency  of  some  organ 
analogous  to  the  thyroid  gland. 

Burger  (1904)  reports  benefit  from  the  use  of  protylin;  Fiirst 
(1904),  from  the  use  of  phytin;  Lepski  (1905)  reported  negative 
results  from  phosphorus  in  cases  complicated  by  severe  general 
illness;  Pfaundler  (1904)  observed  no  difference  in  calcium  ion 
absorption  by  finely  ground  bone  and  cartilage  of  normal  and  rick- 
ety children,  which,  so  far  as  it  goes,  argues  against  the  idea  of  an 
inability  of  rachitic  bones  to  absorb  calcium  salts  in  the  normal 
manner. 

Birk  (1909)  reports  that  phosphorized  cod-liver  oil  increased 
mineral  retention  in  rachitis,  and  that  calcium  retention  varied 
inversely  as  fecal  soap  formation.      (See  table  below.) 


CALCIUM  AND  PHOSPHORUS  METABOLISM- 
RACHITIC   CHILDREN 


-HEALTHY  AND 


Diet 

Condition 

Child 

CaO 

intake 
Grams 

CaO 

retention 
Grams 

Percent 
CaO 
reten- 
tion 

P 

intake 
Grams 

P 

retention 
Grams 

Percent 

P 
reten- 
tion 

Length 

of 
period 

Without  phosphorized 
cod-liver  oil 

Healthy 
Rachitic 

Sch. 
K 
T 
F 

2.175 
2.600 
1.417 
1.629 

+0.942 
+0.515 
+0.021 
-0.135 

43 
20 
2 

4.654 
4.517 

1.862 
2.238 

+0.995 
+0.834 
-0.159 
-0.110 

21 

18 

3  days 
■  • 

With  phosphorized 
cod-liver  oil 

Healthy 
Rachitic 

Sch. 
K 
T 
P 

2.726 
2.390 
1.247 
1.694 

+1.200 
+0.573 
+0.120 
+0.111 

44 
24 
10 

7 

5.716 
4.158 
1.504 
2.386 

+1.951 

+0.708 
-0.131 
+0.201 

32 

17 

'8 

1 1 
•• 

570 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


In  these  data  we  see  no  positive  evidence  of  an  increased  reten- 
tion of  phosphorus,  as  a  result  of  the. administration  of  phosphor- 
ized  cod-liver  oil,  though  there  seems  to  have  been  an  improved 
retention  of  lime. 

Aron  (1908)  found  the  milk  of  the  mothers  of  rachitic  children 
to  be  subnormal  in  lime,  as  also  did  Ramacci  (1910). 

W.  Cronheim  and  E.  Miiller  (1908)  found,  in  balance  experi- 
ments on  normal  and  rickety  children,  that  nitrogen  assimilation 
and  absorption  was  normal  in  the  latter,  and  their  experiments 
showed  nothing  markedly  abnormal  about  the  mineral  balances,  all 
of  which  were  positive. 

Dibbelt  (1908,  1909)  considered,  in  view  of  the  complete- 
ness of  utilization  of  the  calcium  normally  present  in  woman's  milk, 
that  under  abnormal  conditions,  it  is  likely  that  the  calcium  content 
is  often  deficient.  He  also  concluded  that  feeding  with  cow's 
milk  must  often  lead  to  calcium-loss  through  lower  retention, 
especially  through  the  gastrointestinal  disorders  prevalent  at  wean- 
ing time.  Dibbelt  considers  that  the  low  content  of  infants'  food 
in  calcium  phosphate  is  a  predisposing  factor  in  the  development  of 
rickets.  He  puts  much  emphasis  on  gastrointestinal  disorders  as  a 
causative  factor. 

The  following  balances  from  Dibbelt's  work  are  of  interest, 
because,  in  the  light  of  Schabad's  conclusions,  they  may  be  consid- 
ered to  represent  a  case,  not  of  actual,  but  of  pseudorachitis,  since 
there  were  positive  nitrogen  and  phosphorus  balances  coincident 
with  a  marked  loss  of  calcium,  which  last  became  strongly  positive 
simply  through  a  change  of  food. 

DAILY  NITROGEN,   CALCIUM   AND   PHOSPHORUS   BALANCES   WITH 
NORMAL  AND  RACHITIC  INFANTS— Grams 


Subject 


No.  1.    Healthy; 
age  25  days 


No.  2.    Rachitic; 
age  7  months 


No.  2.    Rachitic; 

same  subject  as 

above  but  at  a 

later  date 


Intake 

Outgo 

Retention 

Retention  per  kg-,  body  weight. . 

Intake 

Outgo 

Retention 

Retention  per  kg.  body  weight. . 

Intake  

Outgo 

Retention 

Retention  per  kg.  body  weight.  . 


2.0320 

1.5419 

+0.4901 

+0.1223 


4.1310 

2.7885 
+1.3455 
+0.2790 


5.0400 
3.1E46 

+1.8854 
+0.3840 


CaO 


0.6098 
0.9022 
-0.2924 
-0.0731 


1.1070 

1.4460 

-0.3390 

-0.0704 


1.5030 
0.7495 

+0.7555 
+0.1547 


P2O5 


0.7913 
0.8840 
-0.0933 
-0.0248 


1.4870 

1.4227 

+0.0623 

+0.0123 


1.9058 

1.8256 

+0.0802 

+0.0160 


Diet 


Cow's  milk;  af- 
ter change  from 
mother's  milk 


Specially  pre- 
p  ared  butter- 
milk 


Undiluted  whole 
cow's  milk 


PHOSPHORUS  METABOLISM  571 

Dibbelt  reported  success  in  increasing  the  calcium  content  of 
woman's  milk  from  0.575  to  1.852  parts  per  1000  by  adding  calcium 
salts  to  the  food,  and  that  he  decreased  the  calcium  content  of  dog's 
milk  from  4.56  to  1.94  parts  per  1000  by  use  of  a  ration  low  in 
calcium,  these  results  being  in  marked  contrast  to  the  prevailing 
trend  of  evidence  on  this  matter. 

Bahrdt  and  Edelstein  (1910)  submit  data  showing  the  calcium 
content  of  normal  woman's  milk  to  be  0.0426  percent  CaO,  and  that 
of  the  milk  used  by  rickety  infants  (sustained  by  many  collected 
figures)  as  0.0315  percent  CaO.  Efforts  on  the  part  of  the  authors 
to  increase  the  calcium  content  of  the  milk  of  a  woman  met  with 
apparent  success  in  the  feeding  of  calcium  carbonate,  but  with 
calcium  lactate  the  results  varied  widely,  from  much  below  the 
normal  to  a  little  above. 

Lauxen  (1909)  studied  elemental  phosphorus  administration  to 
young  normal  dogs,  having  in  mind  the  bearing  of  the  results  on 
phosphorus  therapy  in  rachitis.  Phosphorus  was  shown  to  affect  the 
character  of  the  leucocytes  in  a  manner  opposite  to  that  of  rachitis. 
In  the  bones  there  was  a  pronounced  hardening,  with  a  decrease  of 
osteoclasts.  New  bone  formation  was  of  a  type  characteristic  of 
phosphorus  action,  and  decreased  the  marrow  spaces.  Histological 
observation  led  to  the  conclusion  that  phosphorus  has  a  paralyzing 
effect  on  the  bone  cells,  decreasing  both  apposition  and  resorption, 
but  leaving  a  residual  balance  in  favor  of  apposition.  The  spleen 
was  much  enlarged,  possibly  on  account  of  taking  up  blood-forma- 
tion to  compensate  for  restriction  of  this  function  in  the  bone 
marrow. 

Dibbelt  (1909),  in  experiments  on  dogs  with  experimental 
rickets,  concluded  that  human  rickets  is  due  to  anomalies  of  absorp- 
tion and  secretion.  A  late  expression  from  Dibbelt  (1910a)  is  a 
theory  to  the  effect  that  the  cause  of  rickets  is  an  abnormal  amount 
of  calcium-precipitating  compounds,  such  as  phosphorus,  from 
decomposition  of  undigested  casein,  and  carbonates  from  carbohy- 
drate fermentation,  in  the  intestine. 

Flamini  (1907)  submits  calcium  balance  experiments  with  both 
normal  and  rickety  children  under  phosphorus  therapy.  With  the 
former  there  was  a  change  from  a  calcium  retention  of  37.4,  36.S 
and  26.6  percent  of  the  intake  to  37.8,  46.1  and  36.5  percent  after 
12-15  days  of  phosphorus  treatment,  while  with  rickety  children  the 
increase  was  from  59.7,  56.8  and  55.7  percent  to  68.2,  72.6  and 
61.1  percent  after  12-20  days  of  phosphorus  therapy. 


572 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


Gassmann  (1910)  finds  that  lime,  phosphoric  acid  and  carbonic 
acid  all  decrease  in  amount  in  rachitic  bones,  together  with  a  loss  of 
water.  The  relations  Ca:P04:C03  are  about  the  same  for  normal 
and  rachitic  bones.  Magnesium  is  considerably  increased  in  rachitic 
bones. 

ANALYSIS  OF  NORMAL  AND  RACHITIC  BONES— (Gassmann,  1910) 


Aschenheim  and  Kaumheimer  (1911)  find  calcium  and  phos- 
phorus diminished  in  the  muscles  of  rachitic  children,  by  compari- 
son of  their  figures  for  pathological  cases  with  Tobler's  figures  for 
the  normal.  In  6  cases  the  phosphorus  in  the  muscle  varied  from 
0.3849  to  0.6485  percent,  the  normal  being  given  as  0.325  percent. 
In  these  same  six  cases  the  calcium  varied  between  0.0255  and 
0.0570  percent,  while  the  normal  is  given  as  0.0650. 

Schabad  has  conducted  a  most  excellent  series  of  studies  of 
calcium  and  phosphorus  metabolism  in  rachitis  which  contributes 
much  to  our  understanding  of  the  subject.  A  large  part  of  Schabad's 
study  has  centered  in  the  use  of  elemental  phosphorus  in  the  treat- 
ment of  this  disease.  A  comparison  of  its  action  on  rachitic  and 
normal  children  (1907)  .shows  that  in  rickets  phosphorus  assists  in 
the  absorption  and  retention  of  calcium,  and  also  changes  the  com- 
position of  the  bones  toward  the  normal,  but  that  in  health  it 
produces  no  such  effects.  See  also  Schabad  (1908)  and  (1909a). 
Phosphorus  administered  with  cod-liver  oil  has  given  best  results  in 
increasing  retention  of  both  calcium  and  phosphorus,  both  sub- 
stances taking  part  in  the  improvement  (1909b),  while  with  the 
addition  of  calcium  acetate  a  still  larger  retention  of  both  calcium 
and  phosphorus  is  brought  about. 

Calcium  citrate  and  calcium  phosphate  added  to  phosphorized 
cod-liver  oil,  not  only  are  not  retained,  but  they  decrease  the  phos- 
phorus retention.      The  acetate,  however,  as  above  noted,  had  the 


PHOSPHORUS  METABOLISM  573 

opposite  effect,  and  also  brought  about  an  increased  nitrogen  reten- 
tion; though  at  the  same  time  both  the  nitrogen  and  fat  of  the 
feces  were  increased  (1910a,  1910b). 

Lipanin,  olive  oil  and  sesame  oil  cannot  replace  the  cod-liver  oil 
in  the  treatment  of  rickets  (Schabad  and  Sorochowitsch,  1911a), 
and  the  repeated  heating  of  cod-liver  oil  to  100°  for  an  hour  does 
not  destroy  its  beneficial  action  in  rickets  (Schabad  and  Sorocho- 
witsch, 1912). 

Schabad  submits  many  data  to  show  that  only  the  maximum 
normal  calcium  content  of  human  milk  can  cover  the  real  needs  of 
the  infant  for  calcium,  and  that  there  are  doubtless  many  occasions 
when  it  is  insufficient  (1909d).  That  a  deficiency  of  the  milk  in 
calcium  should  be  the  cause  of  rickets  he  considers  not  to  be  exclud- 
ed by  his  studies.  Rickets  is  histologically,  but  not  otherwise, 
distinguishable  from  the  experimental  or  natural  malnutrition  of  the 
bones  of  domestic  animals,  which  Schabad  calls  pseudorachitis.  In 
order  to  identify  a  condition  in  human  beings  parallel  with  the 
pseudorachitis  of  the  domestic  animals  Schabad  assumes  that  both 
actual  and  pseudorachitis  exist  in  human  beings  without  our  being 
able  to  distinguish  them. 

There  is  conflict  in  the  evidence  as  to  whether  or  not  the 
calcium  of  tissues  other  than  bones  is  subnormal  in  rachitis.  Per- 
haps the  difference  in  observations  on  this  point  is  due  to  the  con- 
fusion of  actual  and  pseudorachitis. 

Schabad  finds  the  calcium  content  of  the  milk  of  the  mothers  of 
rickety  children  slightly  lower  than  normal,  and  the  content  of 
organic  nutrients  somewhat  above  normal,  so  that  the  calcium  con- 
tent as  related  to  the  calorific  value  is  considerably  below  the  nor- 
mal. He  finds,  however,  that  rachitis  can  occur  with  a  maximum  cal- 
cium content  of  the  milk,  so  that  it  is  impossible  to  consider  a 
.deficiency  of  the  milk  in  calcium  as  the  only  cause  (1909e,  1911a) 
though  is  has  not  been  so  definitely  related  to  any  other  condition. 
It  has  been  found  impossible  to  increase  the  calcium  content  of 
woman's  milk  by  feeding. 

The  effect  of  rachitis  on  phosphorus  excretion  is  shown  by  the 
following  table  from  Schabad  (1910d)  (page  574). 

For  all  groups  the  phosphorus  excretion  in  rickets  is  distinct- 
ly higher  than  in  health,  and  amounts  in  the  urine  are  smaller.  In 
developing  rickets  calcium  and  phosphorus  excretion  increase 
together,  with  excess  of  the  latter,  and  the  increased  phosphorus 
excretion  usually  exceeds  the  bone-equivalent  of  the  accompanying 
calcium,  so  that  participation  of  other  phosphorus-rich  tissues  is 


574 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


apparent.  The  increase  of  phosphorus  excretion  is  in  the  feces, 
the  urine  phosphorus  decreasing  toward  the  normal,  and  the  normal 
excess  of  urine  phosphorus  over  feces  phosphorus  is  changed  to  an 
excess  of  feces  phosphorus  over  urine  phosphorus.  In  convalescence, 
the  total  phosphorus  excretion  is  subnormal,  and  the  normal  excess 
of  urine  phosphorus  over  feces  phosphorus  returns. 

AVERAGE  DAILY  PHOSPHORUS  EXCRETION  BY  RICKETY  AND 

HEALTHY  CHILDREN  OF  THE  SAME  AGE,  AND  ON  THE 

SAME  DIET 


Diet 

Condition 

P2O5  excretion 
per  kilo  per  day 

Relative  P2O5 
excretion,  in 

percent  of 
amount  sriven 

Partition  of 
P2O5  excre- 
tion 

Excess  in 
feces  in  re- 
lation to 
Ca3(P04)2 

Total 
Grams 

In  urine 
Grams 

Total 

In  urine 

Urine 

Feces 

CaO 

P2O1, 

Breast  fed 

Healthy,  4-5  mos. 
ProgT.  rickets,  5-13  mos. 

0.023 
0.034 

0.018 
0.021 

65.3 
122.2 

52.8 
72.2 

80.6 
60.8 

19.4 
39.2 

64.8 
52.8 

.... 

Infants  on 
cow's  milk 

Healthy,  3-6  mos. 
Progr.  rickets,  5-8M  mos. 

0.214 
0.186 

0.119 
0.077 

79.7 
94.6 

45.1 
39.3 

65.2* 
39.3** 

34.8 
60.7 

22.8 
14.1 

Older  children 
on  mixed  diet 

Healthy,  4-5  yrs. 
Progr.  rickets,  4  yrs. 

0.159 
0.091 

0.102 
0.040 

80.3 
102.1 

51.5 
44.9 

64.4 
44.1 

35.6 
55.9 

22. 

39'.8 

Developed  rickets,  lyr.  5  mos.to  2  yr.  7mos. 

0.178 

0.069 

71.8 

27.5 

38.2 

61.8 

(16.5) 

23.3 

Convalescent,  2 

0.103 

0.077 

65.5 

48.7 

74.4 

25.6 

25.6 

(64) 

*  We  figure  this  to  be  55.6,  and  the  feces  figure  44.4. 

*  *  We  figure  this  to  he  41.4,  and  the  feces  figure  58.6. 

The  calcium  balance  (1910c)  varies  during  the  course  of  a  case 
of  rickets  from  negative  or  subnormally  positive  in  the  progressive 
stage,  to  decided  retention,  which  begins  some  time  before  the  clini- 
cal manifestations  of  improvement,  and  later,  in  convalescence,  to  a 
retention  of  two  or  three  times  the  normal  amount,  which,  on  com- 
plete recovery,  falls  again  to  the  normal. 

Increased  excretion  of  calcium  in  the  progressive  stage  of  the 
, disease  is  through  the  feces;  calcium  excretion  at  this  stage  is- sub- 
normal in  the  urine,  and  in  the  convalescent  stage  above  normal  in 
the  urine.  These  facts,  Schabad  states,  rule  out  the  acid  theory  of 
the  cause  of  rickets. 

Schabad  finds  that  increased  calcium  excretion  in  the  feces 
increases  the  phosphorus  retained  in  the  intestine,  and  conversely 
that  increased  phosphorus  excretion  in  the  feces  increases  the  cal- 
cium retained  in  the  intestine;  therefore,  since  in  rickets  the  feces 
phosphorus  exceeds  the  bone-equivalent  of  the  feces  calcium,  the 
increased  excretion  of  phosphorus,  and  not  of  calcium,  is  the 
primary  factor. 


PHOSPHORUS  METABOLISM 


575 


Schabad  (1909c)  finds  that  in  advanced  stages  of  rickets  the 
relation  of  calcium  to  phosphorus  in  the  bones  changes,  the  calcium 
decreasing,  and  the  phosphorus  increasing,  whereby  the  normal 
relation  of  calcium  to  phosphorus  of  100:75-85  is  changed  to  100: 
70-75,  the  decrease  in  total  ash  affecting  the  calcium  more  than  the 
phosphorus. 

Cattaneo  (1909)  also  notes  departure  from  the  normal  composi- 
tion of  the  bones  in  rachitis,  especially  in  the  direction  of  a  relative 
increase  in  magnesium. 

Rickets  complicated  with  tetany,  Schabad  (1910-11)  finds  not 
to  differ  as  to  calcium  and  phosphorus  metabolism  from  uncompli- 
cated rickets. 

Schabad  states  (1909e)  that  in  pseudorachitis  there  is  no  such 
excess  of  phosphorus  over  calcium  in  the  feces  as  in  true  rickets, 
but  rather,  an  excess  of  calcium  over  phosphorus,  a  fact  which  sug- 
gests a  different  degree  of  participation  of  organs  other  than  the 
bones  in  the  production  of  the  mineral  losses  in  these  diseases. 

As  shown  by  the  table  below,  Ogata  (1911-12)  finds  the  calcium 
phosphate  as  well  as  other  mineral  salts  of  the  bones  much  lowered 
by  rickets.  Different  proportionate  reduction  in  the  amounts  of 
the  several  salts  result  in  an  altered  relation  of  one  to  another  in  the 
bone.      There  was  marked  increase  of  collagen  and  fat. 

ANALYSES  OF  NORMAL  AND  RACHITIC  BONES 
Ogata  (1911-12)— Percent 


Bones  of  2-mos.-old 
healthy  child 

Bones  of  patient  having-  rachitis* 

Tibia 

Ulna 

Femur 

Tibia 

Humerus 

Ribs 

Vertebrae 

65.32 

34.68 

57.54 

1.00 

6.02 

0.73 

33.86 

0.82 

64.07 

35.93 

56.35 

1.00 

6.07 

1.66 

34.92 

1.01 

20.60 

39.40 

14.76 

0.80 

3.00 

1.02 

72.20 

7.20 

33.64 

66.36 

26.94 

0.81 

4.88 

1.08 

60.14 

6.22 

18.88 
81.12 
15.60 

'2i66 

2.62 

81.22 

37.19 
62.91 

32.29 

67.71 

Magnesium  phosphate 

Fat 

*  The  age  of  the  rachitic  patient  was  not  stated. 

Schabad  (1911b,  1911c)  reported  unfavorable  results  from  the 
use  of  the  phosphorized  cod-liver  oil  treatment  in  two  cases  of  so- 
called  late  rickets. 

Schabad  and  Miss  Sorochowitsch  (1911b)  studied  calcium, 
phosphorus,  nitrogen  and  fat  metabolism  in  rachitic  infants  as 
affected  by  various  preparations  of  cod-liver  oil  and  inorganic  salts. 
The  phosphorized  cod-liver  oil  alone  was  found  sometimes  to  be  with- 
out beneficial  effect.  The  most  liberal  calcium  retention  resulted 
from  the  use  of  calcium  acetate  with  cod-liver  oil  emulsion.      Phos- 


576  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

phorus  was  retained  in  considerable  quantity  .from  cod-liver  oil 
emulsion  with  hypophosphites  of  sodium  and  calcium. 

C.  Meyer  (1913)  found  in  metabolism  experiments  with  rickety 
infants  that  phosphorized  cod-liver  oil  caused  a  marked  reduction  in 
the  feces  phosphorus,  and  improved  retention  of  this  element. 

Gessner  (1913)  considers  that  rickets  is  the  result  of  a  dis- 
turbed fat  metabolism,  among  the  causes  of  which  are  deficiency  of 
fat  in  modified  milk,  a  retarded  state  of  metabolism  generally,  and 
the  prevalence  of  acid-reacting  metabolites  in  the  blood  and  lymph. 
He  believes  that  the  efficiency  of  phosphorized  cod-liver  oil  in  the 
treatment  of  rickets  is  due  to  its  ready  emulsification  and  absorp- 
tion, the  presence  of  phosphorus  facilitating  its  oxidation.  Gessner 
likens  this  relation  of  fat  and  phosphorus  to  that  naturally  existing 
in  the  phosphatids  of  the  body. 

Schloss  (1913a,  1913b)  reports  results  from  metabolism  studies 
on  a  breast-fed  infant  under  treatment  for  rachitis.  The  calcium 
balance  was  very  unfavorably  influenced  by  phosphorized  cod-liver 
oil  alone,  but  with  calcium  acetate  added  the  balance  became  and 
continued  to  be  strongly  positive;  the.  phosphorus  balance  was 
affected  much  as  was  that  of  calcium,  though  in  much  lower  degree ; 
a  moderate  retention  of  magnesium  in  the  fore-period  became 
strongly  negative  under  the  influence  of  phosphorized  cod-liver  oil 
and  of  calcium  acetate;  phosphorized  cod-liver  oil  diverted  alkalis 
from  urine  to  feces,  and  the  addition  of  calcium  acetate  led  to  large 
loss  of  alkalis. 

In  a  later  study  Schloss  (1914a)  confirmed  previous  observa- 
tions on  the  ineffectiveness  of  phosphorized  cod-liver  oil  alone,  and 
the  effectiveness  of  the  oil  and  calcium  salts  together  on  the  calcium 
balance.  It  was  found,  further,  that  the  calcium  salt  alone  is 
ineffective,  and  that  two  calcium-casein  preparations  are  of  value  in 
combination  with  phosphorized  cod-liver  oil. 

Regarding  the  phosphorus  metabolism,  the  introduction  of  cal- 
cium acetate  increased  the  feces  phosphorus;  the  addition  of  cod- 
liver  oil  slightly  improved  the  balance,  while  the  oil  alone  caused 
only  a  different  distribution  of  the  phosphorus  elimination.  An 
improvement  of  the  phosphorus  balance  comparable  with  that  of 
calcium  was  first  seen  when  the  calcium  caseonphosphate  and  plas- 
mon  were  used. 

In  a  third  paper  Schloss  (1914b)  reported  once  more  that  phos- 
phorized cod-liver  oil  has  an  unfavorable  influence  on  nitrogen,  cal- 
cium and  phosphorus  metabolism,  and  also  that  CaHP04  with  phos- 
phorized cod-liver  oil  very  greatly  improved  the  calcium  and  especi- 
ally the  phosphorus  balance.  Plasmon  and  CaHP04  were  said  to 
have  identical  effects  on  calcium  and  phosphorus  metabolism. 


PHOSPHORUS  METABOLISM  577 

Kochmann  (1912),  in  balance  experiments  on  dog's,  studied 
the  effect  of  elementary  phosphorus  on  calcium  metabolism.  The 
phosphorus  was  administered  in  quantities  of  1  to  2  mg.  daily.  By 
using*  larger  doses  than  is  possible  in  human  medicine  he  obtained 
positive  results  such  as  have  not  been  obtained  from  the  use  of 
phosphorus  without  cod-liver  oil  with  human  beings.  The  effective 
quantities  equalled  or  exceeded  the  toxic  dose  without  exercising 
an  influence  beyond  the  time  of  administration. 

During  the  use  of  the  phosphorus  there  was  a  regular  and  con- 
tinuous reduction  in  the  preexisting  positive  phosphorus  balance 
until  it  finally  became  a  minus  figure.  The  calcium  balance,  which 
was  negative  during  the  preliminary  feeding,  was  considerably  im- 
proved during  the  phosphorus  therapy,  but  did  not  become  positive 
until  calcium  chloride  and  phosphorus  were  given  together,  which 
produced  marked  retention  of  both  calcium  and  phosphorus.  Mag- 
nesium metabolism  ran  parallel  to  nitrogen  metabolism  and  was  not 
influenced  by  phosphorus  therapy. 

Ribbert  (1913)  reports  a  more  or  less  complete  destruction  of 
the  cartilage  cells  in  rachitis.  From  the  finding  of  these  cell- 
changes  in  light  cases  and  at  the  onset  of  the  disease,  he  concludes 
that  this  condition  is* a  primary  one,  apparently  due  to  some  toxic 
substance  which  hinders  calcium  deposition  in  cartilage  and  bones. 
Ribbert  considers  the  origin  of  this  causative  toxic  substance  to 
lie  in  metabolic  disturbances  resulting  from  wrong  feeding. 

Kassowitz  (1913)  states  his  own  experience  and  also  cites 
results  of  others  showing  that  cod-liver  oil  is  not  essential  to  the 
utilization  of  phosphorus  in  rickets,  that  phosphorus  is  of  value  in 
combination  with  other  oils,  and  of  some  value  alone.  Kassowitz 
cites  cases  in  which  prolonged  treatment  with  cod-liver  oil  alone 
was  not  beneficial.  He  does  not  believe  that  the  calcium  balance 
points  the  way  either  to  the  cause  or  treatment  for  rachitis. 

See  also  Dibbelt  (1913),  Diesing  (1913),  and  Bamberg  and 
Huldschinsky  (1913). 

The  etiology  of  rachitis  remains  an  unsolved  problem. 

THE  THYROID  GLAND  IN  RELATION  TO  PHOSPHORUS  METABOLISM 

The  thyroid  gland  exercises  important  control  over  metabo- 
lism, especially  of  nitrogenous  compounds.  Ingestion  of  thyroid 
gland  substance  increases  tissue  katabolism,  while  deficiency  in  the 
thyroid  causes  cessation  of  physical,  mental  and  sexual  development, 
and  also  retards  ossification  of  epiphyseal  cartilages,  leading  to  the 
characteristic  deformity  of  cretins.  Oxygen  consumption  and  carbon 


578  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL,  5 

dioxide  secretion  are  diminished,  and  metabolism  generally  is  great- 
ly depressed.  There  is  considerable  evidence  that  the  disturbance 
of  functions  is  due  to  toxic  products  of  protein  metabolism,  which, 
through  the  agency  of  the  thyroid  secretion,  are  normally  rendered 
innocuous. 

The  thyroid  stands  in  close  relation  to  the  reproductive  func- 
tions, as  is  shown  by  its  hypertrophy  at  the  time  of  sexual  maturity, 
during  the  menses,  and  during  gestation  and  lactation.  Myxoedema 
and  cretinism  are  associated  with  subnormal  thyroid  secretion; 
while  there  is  some  evidence  that  exophthalmic  goiter  depends  on 
excessive  thyroid  secretion. 

Ord  and  White  (Brit.  Med.  Jour.  1893,  2,  216),  studying 
thyroid  treatment  in  myxoedema,  note  a  slight  increase  in  phos- 
phorus excretion,  though  Houghhardy  and  Langstein  (Jahrb.  fur 
Kinderh.  61,  634,  1905)  observed  a  retention  of  calcium  and  phos- 
phorus, apparently  due  to  the  growth  of  bone  in  a  growing  child. 

Roos  (1895)  found  in  experiments  on  healthy  animals  that  the 
administration  of  thyroid  gland  substance  in  large  doses  caused 
a  marked  increase  in  nitrogen  elimination  (much  more  than  the 
nitrogen  so  introduced)  lasting  for  several  days,  and  also  increased 
sodium  chloride  and  phosphorus  elimination.  The  increased  elim- 
ination of  chlorine  lasted  only  2  or  3  days,  and  then  sank  abruptly  to 
a  markedly  subnormal  amount,  while  the  increased  phosphorus 
elimination,  like  that  of  nitrogen,  persisted. 

The  administration  of  thyroid  substance  to  a  dog,  deprived  of 
the  thyroid  gland,  increased  the  nitrogen  and  chlorine  elimination  to 
a  greater  degree  than  when  given  to  a  normal  animal,  but  the  phos- 
phorus elimination  remained  considerably  below  normal. 

Roos  concludes  that  the  thyroid  gland  has  a  decided  influence 
on  phosphorus  metabolism,  the  nature  of  which  is  as  yet  obscure, 
though  he  notes  evidence  sustaining  the  idea  that  phosphorus  assim- 
ilation requires  the  assistance  of  a  secretory  product  of  the  thyroid. 

Bayon  (1903)  reported  confirmation  of  the  findings  of  others 
in  such  experiments,  that  broken  bones  heal  less  quickly  and  com- 
pletely in  animals  from  which  the  thyroids  have  been  removed,  and 
that  administering  thyroid-gland  preparations  to  such  animals 
favors  the  healing,  though  not  entirely  making  up  for  the  loss  of 
the  glands;  also  that  such  preparations  favor  healing  of  fractures 
in  normal  animals  as  well. 

Scholz  (1905)  has  made  thoroughgoing  studies  of  metabolism 
in  three  cretins,  covering  periods  of  95,  95  and  117  days  respectively, 
and  including  complete  mineral  balances.      The  work  covers  the 


PHOSPHORUS  METABOLISM  579 

normal  state  of  metabolism,  and  the  modifications  produced  by  the 
ingestion  of  thyroid  tablets,  and  of  5  gm.  trisodium  phosphate  per 
day.  Scholz  states  that  metabolism  of  cretins  is  to  be  regarded  as 
sluggish.  The  urine  is  decreased,  and  uric  acid,  creatinin  and  sodium 
chloride  are  excreted  in  subnormal  quantities;  but  urea,  xanthin 
bases,  ammonia  and  sulphur  compounds  appear  to  have  normal 
values.  The  alkaline  earths  show  increased  excretion.  The  phos- 
phorus retention  is  considerable,  on  a  small  intake,  and  does  not 
increase  proportionately  with  increased  intake.  There  was  nothing 
characteristic  in  the  effects  of  thyroid  treatment  on  phosphorus 
metabolism. 

Aeschbacher  (1905-6)  studied  the  thyroid  glands  of  61  persons. 
He  found  the  phosphorus  content  determined  mainly  by  the  richness 
of  the  gland  in  nuclei,  but  in  part  by  the  phosphorus  content  of  the 
colloid.  As  noted  by  Kocher,  there  was  generally  a  reciprocal  rela- 
tion between  the  quantities  of  phosphorus  and  iodine  in  the  gland, 
due  principally  to  the  fact  that  glands  rich  in  the  iodine-containing 
colloid  are  relatively  poor  in  cells.  The  thyroids  of  women  were 
found  larger  and  also  richer  in  iodine,  than  the  thyroids  of  men, 
while  the  phosphorus  value  reaches  the  higher  figure  in  the  thyroids 
of  men.  Diseases  which  cause  circulatory  disturbances  cause  a 
marked  decrease  in  the  iodine  content  of  the  gland,  but  for  phospho- 
rus this  relation  does  not  hold. 

Saccone  (1907)  removed  the  thyroids  and  parathyroids  from  a 
dog,  after  which  the  urinary  phosphorus  rose  to  three  times  the 
previous  amount.  The  feeding  of  a  phosphatic  diet  containing  liver, 
brain  and  sodium  glycerophosphate  did  not  bring  about  normal  con- 
ditions, but  instead,  increased  especially  the  organic  phosphorus  of 
the  urine ;  the  feeding  of  the  normal  diet,  however,  with  the  addition 
of  thyroidin  reduced  the  excretion  of  phosphorus  to  the  original 
level. 

Underhill  and  Saiki  (1908)  in  the  few  urinary  phosphorus 
estimations  made  in  their  metabolism  study  note  a  low  phosphorus 
elimination  "possibly  indicating  when  considered  with  the  low  purin 
and  allantoin  excretion,  a  low  rate  of  nuclear  disintegration.  A 
large  output  of  purin-nitrogen  and  a  low  output  of  phosphorus  were 
observed  after  the  continued  administration  of  large  doses  of 
thyroid  tissue." 

Soli  (1909)  finds  that  rabbits,  if  deprived  of  thyroids  at  a  suf- 
ficiently early  age,  fail  to  develop  the  skeleton  normally,  while 
guinea  pigs  and  chickens  did  not  show  this  result  so  readily. 


680  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Bircher  (1910)  removed  the  thyroid  glands  from  6  rats  of  the 
same  litter.  To  three  of  these  he  fed  thyroidin  tablets.  The 
others  were  used  as  controls.  The  former  showed  much  the  more 
rapid  calcification  of  the  bones. 

A.  Juschtschenko  (1911)  observed  that  thyroidectomy  reduced 
the  nuclease  content  of  young  dogs. 

Schafer  (1912)  found  that  the  addition  of  small  amounts  of 
thyroid  tissue  to  the  diet  of  white  rats  greatly  increased  the  food 
consumption,  especially  in  quite  young  individuals,  with  acceleration 
of  growth,  and  retention  of  nitrogen.  Phosphorus  excretion  is 
increased  if  the  ingestion  of  thyroid  is  increased  beyond  a  certain 
point. 

A.  I.  Ushenko  (1913)  found  that  after  thyroidectomy  the  ratio 
of  phosphorus  to  nitrogen  in  the  urine  first  increased  and  then 
decreased,  synthetic  processes  suffering  acute  disturbance  in  tissues 
containing  nitrogen  and  phosphorus. 

A.  S.  Juschtschenko  (1913)  found  in  experiments  on  dogs  that 
thyroidectomy  causes  a  general  disturbance  of  phosphorus  com- 
pounds in  the  organs  and  tissues.  There  was  general  increase  of 
the  inorganic  phosphorus  of  brain,  heart,  pancreas  and  liver,  and 
general  decrease  of  the  total  and  organic  phosphorus  of  the  same. 
In  the  kidneys  there  was  increase  of  the  total  phosphorus,  generally 
both  organic  and  inorganic;  and  in  the  serum  organic  and  total 
phosphorus  increased,  while  inorganic  decreased. 

Hyperthyroidism  was  also  found  to  cause  disturbance  in  the 
phosphorus  distribution  in  the  tissues,  these  disturbances  being  in 
some  respects  just  the  opposite  to  those  observed  in  athyroidism. 
The  organic  and  total  phosphorus  are  decreased  in  brain,  muscle 
and  heart,  but  are  increased  in  liver,  kidney,  pancreas  and  serum. 
Inorganic  phosphorus  was  less  in  all  the  tissues  than  in  the  normal 
animal,  a  fact  almost  completely  opposite  to  the  results  in  thyroid- 
ectomy. 

Confusion  has  resulted,  in  the  study  of  the  function  of  the  thy- 
roid, through  the  removal  along  with  the  thyroid  of  the  parathy- 
roids, or  parts  of  the  same,  or  through  their  injury.  Further  study 
will  more  sharply  differentiate  their  functions. 

Karl  Droge  (1913)  has  recently  reported  experiments  with 
young  dogs  in  which  observations  were  made  on  the  effects  during 
the  suckling  period  of  removal  of  thyroids,  spleen  or  testicles.  Six 
pups  of  a  litter  of  eight  were  used.  Dog  I  was  killed  about  12  hours 
after  birth ;  dog  II  was  kept  as  a  control ;  the  thyroid  dogs  I  and  II, 
the  spleen-dog,  and  the  testicle-dog  were  operated  upon  on  the  day 


PHOSPHORUS  METABOLISM 


581 


the  weight  had  about  doubled,  the  10th  day ;  ail  were  suckled  by  the 
mother.  On  the  23rd  day  the  thyroid-dog  I  was  killed  to  allow  better 
nourishment  of  the  others ;  the  remaining  dogs  were  killed  as  soon 
as  there  was  any  tendency  shown  to  take  other  food  than  the 
mother's  milk,  the  28th  day.  The  bodies  were  analyzed  as  a  whole 
for  moisture,  fat  and  ash,  and  for  nitrogen,  calcium,  magnesium  and 
phosphorus  in  the  fat-free  dry  substance  and  in  the  ether  extract. 
We  quote  but  two  of  the  44  tables  of  results  given.  Conclusions 
should  await  results  with  more  individuals. 


LIME,  MAGNESIUM  AND  PHOSPHORUS  CONTENT  IN  RELATION  TO 
NITROGEN  (Droge,  1913) 


Day  of 
operation 

Day 
killed 

Nitrogen 
content 

Gm. 

To  1  gm.  nitrogen 

Dog 

CaO 
Gm. 

MgO 
Gm. 

P2O5 
Gm. 

Dog  I 

iotii 

1st 
28th 
23rd 
28th 
ii 

6.87 
.    30.63 
21.85 
17.06 
19.48 
35.34 

0.31 
0.49 
0.56 
0.44 
0.74 
0.52 

0.003 
0.008 
0.008 
0.008 
0.004 
0.005 

0.43 

Dog-  II ... 

0.60 

0.46 

0.62 

0.67 

0.54 

PHOSPHORUS  CONTENT  IN  FAT-FREE  DRY  SUBSTANCE  AND  IN 
ETHER-SOLUBLE  SUBSTANCE   (Droge,  1913) 


P2O5  in  fat-free  dry  substance 

P2O5  in  ether-soluble  substance 

Dog 

Weight  of 
fat-free  dry 
substance 

Grams 

P2O5 
Percent 

P2O5 

Grams 

Weight  of    ! 
ether-soluble 
substance 

Grams 

P2O5 
Percent 

P2O5 
Grams 

Total 
P2O5 

Grams 

Dog  I 

61.1372 

277.70 
193.72 
146.27 
182.51 
324.70 

4.85 
6.51 
5.09 
7.16 
7.01 
5.81 

2.965 
18.078 

9.860 
10.473 
12.794 
18.865 

10.6278 
280.507 
208.570 
255.656 
178.810 
312.977 

? 
0.09 
0.14 
0.045 
0.11 
0.07 

? 

0.252 
0.292 
0.115 
0.197 
0.219 

2.965 

Dog  II 

18.330 

Thyroid-dog  I 

10.152 

10.588 
12.991 

19.084 

MORBUS  BASEDOWII— EXOPHTHALMIC  GOITER 

Scholz  (1895)  conducted  a  complete  balance  experiment  with  a 
woman  suffering  from  exophthalmic  goiter,  and  with  a  healthy 
person  on  the  same  treatment  for  comparison,  two  periods  of  3-5 
days  with  each  subject,  one  without  treatment,  and  the  other  with 
dried  thyroid  gland  substance. 

Without  treatment  the  goiter  patient  stored  7.437  gm.  nitrogen 
and  1.06  gm.  P,05  per  day;  with  thyroid  treatment  the  nitrogen 
storage  was  7.09  gm.  per  day,  while  the  P205  balance  fell  from  +1.06 
to— 2.09  gm. 


582  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

Under  thyroid  treatment  the  nitrogen  storage  with  the  healthy 
subject  fell  from  +3.76  to  +2.64  gm.,  while  the  P205  balance  fell 
from  —2.996  to  —3.831  gm. 

There  was  not  any  great  increase  of  phosphorus  elimination  in 
the  urine  with  either  subject,  but  in  the  feces  the  phosphorus  elim- 
ination of  the  goiter  patient  was  increased  10-fold,  and  that  of  the 
healthy  subject  about  25  percent.  Since  there  was  no  such  increase 
of  nitrogen  outgo  as  of  phosphorus  it  was  evident  that  the  latter 
was  due  to  phosphate  rather  than  nuclein  katabolism. 

Von  Noorden  mentions  the  similar  result  of  thyroid  treatment 
on  nitrogen  metabolism  by  Hirschlaff  (Zeit.  klin.  Med.  36,  200, 
1899),  and,  on  the  other  hand,  the  greatly  increased  excretion  of 
nitrogen  as  observed  by  Matthes  (Verhandl.  der  Cong.  f.  inn.  Med. 
(1897),  232)  and  by  David  (Zeit.  Heilkunde  17,  439). 

That  the  behavior  of  Scholz's  patient  under  thyroid  treatment 
was  typical  seems  not  yet  to  have  been  demonstrated,  though  Roos 
(1895)  and  others  obtained  similar  results,  though  less  marked,  in 
experiments  with  animals. 

Berkley  (1908)  is  said  to  have  administered  an  alcoholic  solu- 
tion of  lecithin  in  cases  of  exophthalmic  goiter  with  strikingly  favor- 
able results.  Berkley  used  this  solution  for  alternate  weeks  with 
glycerophosphates,  quinine  and  gentian,  but  with  unfavorable 
results  from  the  glycerophosphates.  With  lecithin  there  was  gain 
in  weight  and  relief  from  nervous  symptoms,  while  with  glycero- 
phosphates there  was  loss  in  weight,  and  increase  of  nervous 
phenomena.  Lecithin  treatment  was  stated  to  be  out  of  place  with  a 
disturbed  digestion,  and  not  of  value  without  the  assistance  of  a 
milk  diet. 

Tschikste  (1911)  considers,  as  a  result  of  a  metabolism  study, 
that  the  iodin-free  nucleoprotein  which  Oswald  isolated  from  the 
thyroid  has  an  influence  in  exophthalmic  goiter  opposite  to  that  of 
thyreoidin  (iodothyrin) .  In  this  article  Tschikste  reviews  many 
articles  on  this  subject  that  we  have  not  consulted. 

TOXINS,   ANTITOXINS   AND   PHOSPHORUS   METABOLISM 

Dalmastri  (1901)  studied  phosphorus  metabolism  as  affected 
by  rabes  antitoxin.  Metabolism  generally  was  considerably 
increased  during  the  cure,  as  shown  by  an  increased  loss  of  nitro- 
gen and  especially  of  phosphorus,  and  by  loss  of  weight.  The 
period  following  showed  a  prompt  return  to  normal,  so  far  as  nitro- 
gen and  body  weight  go,  but  a  slow  return  as  to  phosphorus. 


PHOSPHORUS  METABOLISM  583 

Dmitriewski  (1900)  determined  the  effects  on  metabolism  in 
the  dog  of  poisonous  substances  extracted  from  Bacillus  pyocyaneus 
and  Bacillus  coli  communis,  and  also  of  diphtheria  toxin.  In  carefully 
controlled  subcutaneous  injection  experiments,  it  was  found  that  all 
three  of  these  poisons  led  to  increased  nitrogen  and  corresponding 
phosphorus  elimination. 

Novi  (1904)  published  balance  data  on  phosphorus  metabolism 
during  antirabes  treatment,  both  with  the  virulent  and  non-virulent 
material,  and  also  with  and  without  sodium  glycerophosphate,  the 
latter  being  administered  in  one  period  per  os,  and  in  another  by  in- 
jection. Beyond  a  slight  increase  of  urinary  phosphorus,  under  the 
influence  of  the  antirabes  treatment,  the  figures  did  not  show  any 
notable  departures  from  the  normal,  though  the  author  states  that 
the  cure  by  both  the  toxic  and  the  non-toxic  treatment  caused  a 
leucocytosis  and  a  following  or  coexistent  leucolysis.  See  also 
Decroly  (1898). 

Lederer  and  Stolte  (1911)  made  mineral  analyses  of  human  and 
of  dog  hearts  under  normal  conditions  and  under  the  influence  of 
diphtheria  or  scarlet  fever  toxins.  The  authors  did  not  find  that 
these  diseases  alter  the  composition  of  the  heart. 

TUBERCULOSIS 

The  effects  of  tuberculosis  on  phosphorus  metabolism  are  of 
several  kinds;  (1)  characteristic  changes,  both  quantitative  and 
qualitative,  in  the  phosphorus-containing  lipoids  of  the  tissues,  (2) 
pathologic  retention  and  deposit  of  phosphates  in  the  calcification 
of  degenerative  tissues,  (3)  deranged  organic  functions  of  the  parts 
affected,  which  of  course  may  be  of  most  diverse  character,  (4) 
others  due  to  the  fever  induced  by  this  disease,  (5)  still  others  due 
to  anaemia,  and  lastly,  (6)  those  of  general  starvation.  The  relation 
of  tuberculosis  to  phosphorus  metabolism,  therefore,  is  largely  gen- 
eral and  incidental,  and  little  attention  has  been  given  to  the  matter, 
except  in  relation  to  diet.  In  times  past  much  prominence  has 
been  given  the  idea  of  flooding  the  system  with  lecithin,  as  in  the 
very  liberal  ingestion  of  eggs,  and  benefit  derived  from  such  treat- 
ment has  often  been  ascribed  in  large  part  to  the  phosphorus  com- 
pounds present.  Lecithin  and  nuclein  phosphorus  have  also  had 
a  place  in  the  treatment  of  this  disease  by  parenteral  injection.  At 
present,  however,  an  appreciation  of  the  greater  importance  of  the 
environment  and  general  management  has  detracted  emphasis  from 
this  particular  dietetic  consideration. 


584  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

'  PHOSPHORUS  METABOLISM  IN  TUBERCULOSIS 

Passing  over  the  earlier  work  referring  to  this  matter,  we  note 
Mitulesco's  (1902)  observation  that  in  times  of  hemorrhage  from 
the  lungs  in  tuberculosis  there  is  a  decrease  of  urinary  nitrogen  and 
phosphorus.  In  a  later  article  (1903a),  he  reports  complete  nitro- 
gen and  phosphorus  balances  on  four  cases  of  tuberculosis.  Three 
were  given  tuberculin  treatment;  the  fourth  hygienic  and  dietetic 
treatment  only.  The  last  mentioned  patient  improved  slowly,  and 
the  positive  nitrogen  and  phosphorus  balances  were  increased.  The 
three  patients  on  tuberculin  treatment  were  further  advanced  in  the 
disease,  all  showing  negative  nitrogen  and  phosphorus  balances. 
Under  this  treatment  these  balances  all  became  positive,  the 
improvement  being  greater  than  with  the  other  case. 

In  another  paper  Mitulesco  (1903b)  reports  nitrogen  and 
phosphorus  balances  on  eight  cases  of  tuberculosis.  In  general  the 
loss  of  nitrogen  and  phosphorus  in  the  urine  was  excessive,  and  the 
balances  negative.  There  was  a  high  content  of  organic  phosphorus 
in  the  urine.  The  nitrogen  and  phosphorus  in  the  urine  were  less 
at  times  of  hemorrhage.  The  loss  of  appetite,  impairment  of  digest- 
ive functions,  increased  cell  destruction,  the  resultant  anaemia,  and 
the  excessive  losses  of  material  by  the  body  combine  to  lower  the 
resistance  of  the  organism  to  the  infection. 

Zickgraf  (1910)  compares  phosphorus  and  chlorine  elimination 
in  tuberculosis  and  chlorosis.  He  finds  the  average  phosphorus 
excretion  to  be,  as  Teissier  stated,  lower  in  chlorosis  than  in  tuber- 
culosis ;  but  the  difference  is  too  small,  and  the  individual  variations 
too  great,  to  make  possible  diagnostic  use  of  the  distinction. 

Mulier  (1911)  concludes  that  the  phosphorus  excretion  in  tuber- 
culosis is  not  characteristic,  and  has  no  differential  diagnostic  value. 

DEMINERALIZATION  IN  TUBERCULOSIS 

Ott  (1903)  reports  complete  mineral  and  nitrogen  balances 
with  three  cases  of  tuberculosis.  He  admits  the  presence  of  "demin- 
eralization"  in  certain  cases,  but  finds  that  it  is  not  a  regular  symp- 
tom. 

Steinitz  and  Weigert  (1904, 1905)  analyzed  the  body  of  a  tuber- 
culous child,  and  compared  the  analysis  with  others  by  Sommerfeld 
on  a  child  dead  from  gastrointestinal  disorder,  another  such  by 
Steinitz,  and  six  analyses  of  new-born  infants  by  Camerer  and 
Soldner.  They  find  in  their  study  no  grounds  for  belief  in  the 
theory  of  demineralization. 


PHOSPHORUS  METABOLISM  585 

Amat  (1906),  also  studying  demineralization,  considered  the 
possible  diminution  of  the  phosphorus  content  of  the  body  through 
the  use  of  white  flour  as  a  predisposing  factor  to  tuberculous  infec- 
tion. In,  this 'study  mice  were  fed  on  breads  made  from  white 
roller  process  flour,  and  flour  ground  between  stones.  The  latter 
was  eaten  in  larger  quantity  and  produced  a  considerable  gain  in 
weight ;  the  former  (the  roller  process  flour)  was  not  eaten  in  quite 
so  great  quantities,  and  did  not  maintain  the  body  weight.  No  evi- 
dence was  submitted  on  demineralization. 

A.  Mayer  (1907)  studied  metabolism  in  tuberculosis,  and  from 
complete  nitrogen  and  mineral  balances  with  five  patients  in  3-day 
periods  draws  conclusions  as  to  demineralization.  In  these  cases 
he  finds  a  retention  of  calcium  and  phosphorus  and  a  marked  reten- 
tion of  chlorine.  There  was  a  slight  tendency  to  retain  potassium 
and  a  greater  one  to  retain  sodium.  There  was  no  apparent  rela- 
tion between  the  phosphorus  and  nitrogen  balances.  Mayer's  posi- 
tive mineral  balances  lead  him  to  the  belief  that  demineralization  is 
not  characteristic  of  tuberculosis. 

Sarvonat  and  Rebattu  (1910)  made  ash,  calcium  and  phospho- 
rus estimations  on  normal  and  tuberculous  guinea  pigs.  They  con- 
clude that  the  total  ash  of  the  body  of  the  tuberculous  guinea  pig 
decreases  about  one-tenth,  this  loss  being  due  to  the  animal's  grow- 
ing thin  in  flesh.  The  absolute  amount  of  phosphorus  was  less  in  the 
body  of  the  tuberculous  animals,  and  the  proportion  of  phosphorus 
to  total  ash  was  much  less,  while  the  authors  state  that  the  phos- 
phorus in  the  ash  of  the  soft  parts  was  sometimes  the  higher  in  the 
tuberculous  animals.  The  absolute  and  relative  calcium  content  of 
the  bones  of  the  tuberculous  animals  is  decreased.  The  propor- 
tionate loss  of  calcium  from  the  bones  is  much  greater  than  from  the 
soft  parts.  In  these  cases,  then,  there  was  demineralization — as,  in 
fact,  there  is  in  starvation  from  whatever  cause. 

PHOSPHORUS  COMPOUND  THEBAPY  IN  TUBERCULOSIS 

Gilbert  (1901)  states  that  consumptives  and  neurasthenics 
thrive  remarkably  under  the  influence  of  pills  containing  0.1  to  0.5 
gm.  lecithin.  Lecithin  was  also  administered  subcutaneously  in 
injections  of  0.05  to  0.15  gm.,  repeated  every  second  day.  The 
appetite  and  weight  increased,  and  no  evil  effects  were  observed, 
even  on  prolonged  administration.  One  subject,  already  in  a  hos- 
pital for  four  months,  gained  3.5  kg.  in  one  month.  Other  details 
of  improvement  were  noted. 


586  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

According  to  Morichau-Beauchant  (1901)  the  efficacy  of  leci- 
thin therapy  in  tuberculosis  is  inversely  as  the  stage  of  advancement 
of  the  disease. 

Gilbert  and  Fournier  (1901)  report  that  egg  lecithin  in  doses 
of  0.10  to  0.50  gm.  per  day  by  the  mouth,  or  0.05  to  0.15  gm.  every 
other  day  by  injection,  led  to  general  improvement,  and  gain  in 
appetite,  and  in  weight,  in  cases  of  pulmonary  tuberculosis. 

Claude  and  Zaky  (1901a)  found  by  feeding  and  injection 
experiments  with  lecithin,  on  tuberculous  men  and  guinea  pigs, 
increased  urinary  nitrogen  and  decreased  urinary  phosphorus, 
associated  with  gain  in  weight  and  general  improvement.  The  leci- 
thin did  not  affect  the  development  of  the  disease.  The  improve- 
ment was  marked  in  early  stages  of  the  disease,  but  perceptible  even 
in  those  which  were  hopelessly  advanced.  See  also  (1901b)  and 
(1901c). 

Mouneyrat  (1902a)  experimented  with  disodium-methyl-arsen- 
ate  and  nucleic  acid  for  tuberculosis.  He  reports  results  with  120 
patients.  The  daily  dose  was  30  c.c.  of  a  solution,  taken  in  two 
portions,  the  whole  containing  0.05  gm.  methyl  arsenate  of  sodium 
and  0.20  gm.  nucleic  acid  from  herring  milt.  Both  red  and  white 
blood  corpuscles  increased  rapidly,  and  general  improvement  was 
marked  after  varying  periods  of  treatment  up  to  a  maximum  of  a 
month,  the  Koch  bacillus  disappearing  from  the  spittle  in  many 
cases.  This  treatment  is  said  to  have  succeeded  where  lecithin 
treatment  failed. 

Colombet  (1902)  administered  this  same  arsenic-phosphorus 
compound  with  33  cases  of  chronic  tuberculosis.  Two  cases  died; 
3  did  not  receive  benefit ;  the  remainder  were  said  to  be  improved  in 
a  notable  manner,  especially  those  having  the  disease  in  the  first 
or  second  degree.  Many  symptoms  of  improvement  were  noted, 
among  them  diminution  of  phosphaturia. 

Ward  (1910)  attempted  to  relieve  the  progressively  increasing 
anaemia  of  tuberculosis  by  intravenous  injection  of  nuclein-saline 
solution.  This  treatment  caused  a  rapid  disappearance  of  the  poikilo- 
cytes,  and  their  replacement  by  new  and  healthy  erythrocytes.  The 
data  presented  show  that  the  nuclein  treatment  markedly  increased, 
nearly  to  the  normal,  the  haemoglobin,  the  number  of  red  cells,  and 
the  specific  gravity  of  the  blood.  Of  the  15  cases  reported,  2  died, 
9  recovered  and  became  free  from  tubercle  bacilli,  4  were  improved 
but  not  yet  cured,  while  one  refused  the  treatment  after  the  firit 
period.      The  solution  used  was  composed  as  follows : 


PHOSPHORUS  METABOLISM  587 

Approximately  6  grains  of  sodium  triticonucleate  were  dissolved 
in  each  ounce  of  physiological  salt  solution,  and  standardized  to  1 
mg.  organic  phosphorus  to  each  cubic  centimeter.  This  solution 
was  used  in  the  quantity  of  1  oz.  per  20  lbs.  body  weight  of  patient. 

Otolski  and  Biernacki  (1912)  made  total  phosphorus  and  phos- 
phatid  phosphorus  determinations  on  the  organs  of  rabbits  vaccin- 
ated with  dead  tubercle  bacilli,  and  compared  these  with  similar 
determinations  on  the  organs  of  a  control  rabbit.  The  data  reported 
show  unmistakably  lower  content  in  both  these  items  in  the  kidneys, 
heart  and  lungs  of  the  treated  animals  than  in  the  control;  in  the 
livers  it  was  lower  in  some  cases  but  higher  in  most.  The  results 
showed  a  loss  of  lecithins  and  an  increase  of  jecorins  in  the  liver, 
and  at  the  same  time  a  marked  increase  in  the  phosphorus  content 
of  the  lecithin. 

Griniew  (1913)  has  made  an  investigation  of  the  quantitative 
differences  in  the  lipoids  and  the  lipoid  phosphorus  of  several 
organs  of  guinea  pigs,  brought  about  by  chronic  tuberculosis.  The 
organs  were  extracted  consecutively  with  acetone,  benzol,  petroleum 
ether,  alcohol  and  ethyl  ether,  and  the  amounts  of  the  extracts  and 
of  their  phosphorus  content  were  recorded.  Full  data  are  given  and 
compared  in  tables.  We  quote  three  condensed  tables  (p.  588)  and 
a  part  of  the  author's  conclusions. 

Conclusions:  "During  tuberculous  infection  the  chemical 
constitution  of  the  cells  of  nearly  all  organs  and  tissues  changes  in 
its  lipoidal  part.  This  change  is  of  qualitative  as  well  as  quantita- 
tive character.  It  shows  itself  in  the  diminution  of  the  quantity  of 
phosphorus  in  the  lipoids  and  by  the  replacement  of  different  forms 
of  lipoids  by  each  other 

"During  tuberculosis:  (a)  In  nearly  all  the  organs  the  sum  of 
all  the  lipoids  and  the  phosphorus  diminishes  as  compared  with  the 
normal.  (b)  The  quantity  of  cholesterin  is  increased  in  certain 
organs,  diminished  in  others,  (c)  In  all  the  organs  there  is  less  of  the 
extracts  containing  lecithin,  (d)  In  all  the  organs  there  is  more  of 
the  extracts  containing  cephalin  and  myelinous  materials,  (e)  In  all 
the  organs  the  diminution  of  the  quantity  of  lipoidal  material  of  the 
filtrate  from  the  acetone  and  of  the  phosphorus  contained  in  it  is 
pronounced,  (f )  The  quantity  of  materials  extracted  by  benzol  is 
comparatively  increased  in  most  of  the  organs,  (g)  It  appears  that 
a  part  of  the  lecithin  passes  into  cephalin  or  analogous  compounds, 
(h)  The  organs  which  suffer  most  in  their  lipoid  content  and  in 
phosphorus  contained  there  are :  the  lungs,  the  spleen,  the  medulla, 
the  liver.      The  lungs  suffer  most." 


588 


OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 


LIPOIDS  OF  THE  ORGANS  OF  NORMAL  GUINEA  PIGS 
Griniew  (1913) — Percent,  Dry  Basis 


Mixture  of 

Lecithin 

Cephalin 

Phosphatids 

Lipoids  of 

the  lipoids 

of  benzol 
extract  +Cd 

the  alcohol 
extract  solu- 

of the 
acetone  and 

Phos- 

Phos-l 

compounds 

ble  in  water 

alcohol  ex- 

tissues 

phorus 

phatids 

Ex- 
tract 

Ex- 
tract 

tracts 

P 

P 

Ex- 
tract 

P 

Ex- 
tract 

P 

Ex- 
tract 

P 

Liver 

0.3 

15.9 

1.2 

0.022 

0.5 

0.011 

2.7 

0.03 

2.04 

0.03 

9.5 

0.18 

Kidneys 

0.2 

14.1 

1.0 

0.023 

0.34 

0.008 

3.7 

0.015 

4.1 

0.06 

5.0 

0.09 

Brain 

0.6 

31.8 

0.5 

0.007 

2.5 

0.05 

2.05 

0.4 

5.4 

0.07 

2.8 

0.06 

Heart 

0.15 

12.2 

0.73 

0.018 

0.31 

0.006 

1.65 

0.003 

3.7 

0.05 

5.7 

0.08 

Muscles 

0.16 

7.3 

0.9 

0.016 

0.35 

0.008 

1.8 

0.008 

2.02 

0.02 

2.2 

0.03 

Lung-s 

0.2 

9.7 

1.56 

0.03 

1.1 

0.038 

1.83 

0.006 

2.3 

0.05 

2.8 

0.07 

Spleen 

0.25 

14.4 

0.3 

0.04 

0.6 

0.055 

2.08 

0.0002 

7.5 

0.12 

3.8 

0.07 

Medulla 

0.04 

1.9 

0.15 

0.003 

0.17 

0.004 

0.43 

0.43 

0.004 

0.7 

0.01 

(x)      Apparently  the  heading   "Lipoids"   given  here  in  the  original  is  a  mistake. 

LIPOIDS  OF  THE  ORGANS  OF  TUBERCULOUS  GUINEA  PIGS 
Griniew  (1913) — Percent,  Dry  Basis 


Orgrans 

and 
tissues 

Phos- 
phorus 

Phos- 
phatids 

Lecithin 

Cephalin, 

myelin,  and 

others 

Phosphatids 
of  benzol  ex- 
tract and  of 
the  Cd  com- 
pounds 

Mixture  of  the 
lipoids  of  the 
filtrate  from 
the  acetone 

The  portion 
of  the  alco- 
hol extract 
soluble  in 
water 

Ex- 
tract 

P 

Ex- 
tract 

P 

Ex- 
tract 

P 

Ex- 
tract 

P 

Ex- 
tract 

P 

Liver 

Kidneys 

Brain 

Heart 

Muscles 

Lungs 

Spleen 

Medulla 

0.13 
0.09 
0.56 
0.15 
0.08 
0.14 
0.14 
0.02 

9.5 
13.7 
28.6 
11.7 

9.4 
10.0 
11.5 

2.2 

0.58 
0.97 
0.42 
0.65 
0.59 
0.37 
0.16 
0.08 

0.011 

0.03 

0.012 

0.02 

0.011 

0.011 

0.003 

0.0009 

0.8 

0.73 

4.7 

1.02 

0.30 

0.4 

0.6 

0.12 

0.022 

0.0093 

0.072 

0.02 

0.0084 

0.007 

0.006 

0.0022 

1.9 
7.1 
16.9 
3.1 
3.5 
2.1 
2.4 
0.83 

0.01 

0.02 

0.4 

0.007 

0.01 

0.015 

6.005 

3.4 
0.3 
2.6 
2.9 
0.8 
3.1 
0.8 
0.04 

0.05 

0.004 

0.04 

0.04 

0.01 

0.03 

0.006 

0.0005 

2.6 
4.6 
3.8 
3.9 
4.1 
3.2 
7.5 
1.1 

0.04 

0.02 

0.05 

0.06 

0.04 

0.07 

0.1 

0.01 

COMPARISON  OF  LIPOIDS  AND  PHOSPHORUS  IN  NORMAL  AND 
TUBERCULOUS  ORGANS— (Griniew,  1913)  Percent,  Dry  Basis 


Organs 
and 

tissues 


Liver  . . . 
Kidneys 
Brain... 
Heart..  . 
Muscles. 
Lung-s.. . 
Spleen... 
Medulla. 


Phosphatids 


Nor- 
mal 


15.9 
14.1 
31.8 
12.2 
7.3 
9.7 
14.4 
1.9 


Pathol. 


9.5 
13.7 
28.6 
11.7 

9.4 
10.0 
11.5 

2.2 


Differ- 
ence 


-6.4 
-0.4 
-3.2 
-0.5 
+2.1 
4-0.3 
-2.9 
+0.3 


Nor- 
mal 


26.6 
32.1 
47.9 
19.5 
16.8 
21.1 
29.8 
8.7 


Lipoids 


Pathol. 


21.5 
34.8 
50.4 
19.1 
13.6 
19.7 
22.4 
4.9 


Differ- 
ence 


-5.1 
+2.7 
+2.5 
-0.4 
-3.2 
-1.4 
-7.4 
-3.8 


Nor- 
mal 


0.30 
0.20 
0.60 
0.15 
0.16 
0.20 
0.25 
0.04 


Phosphorus 


Pathol. 


0.13 
0.09 
0.56 
0.15 
0.08 
0.14 
0.14 
0.02 


Differ- 
ence 


-0.17 
-0.11 
-0.04 

0 
-0.08 
-0.06 
-0.11 
-0.02 


PHOSPHORUS  METABOLISM  689 


BIBLIOGRAPHY 


1897  Abderhalden,  Emil:  Zur  quantitativen  Analyse  des  Blutes,  Zeit.  physiol. 

Chem.,  23,  521-531. 

1898  Abderhalden,  Emil:  Zur  quantitativen  vergleichenden  Analyse  des  Blutes, 

Ibid.,  25,  65-115. 
1899a  Abderhalden,  Emil:  Die  Beziehungen  der    Wachsthumsgeschwindigkeit 

des  Sauglings  zur  Zusammensetzung  der  Milch  beim  Kaninchen,  bei 

der  Katze  und  beim  Hunde,  Ibid.,  26,  487-497. 
1899b  Abderhalden,  Emil:  Die  Beziehungen  der  Zusammensetzung  der  Asche 

des  Sauglings  zu  derjenigen  der  Asche  der  Milch,  Ibid.,  26,  498-500. 
1899c  Abderhalden,  Emil:  Die  Beziehungen  der   Zusammensetzung  der  Asche 

des  Sauglings  zu  derjenigen  der  Asche  der  Milch  beim  Meerschwein- 

chen,  Ibid.,  27,  356-367. 
1899d  Abderhalden,  Emil:  Die  Beziehungen    der    Wachsthumsgeschwindigkeit 

des  Sauglings  zur  Zusammensetzung  der  Milch  beim  Hunde,  beim 

Schwein,  beim  Schaf,  bei  der  Ziege  und  beim  Meerschweinchen,  Ibid., 

27,  408-462;  594. 

1905  Abderhalden,  Emil:  Abbau  und  Aufbau  der  Eiweisskorper  im  tierischen 

Organismus,  Ibid.,  44,  17-52. 
1912    Abderhalden,  Emil:    Futterungsversuche    mit    vollstandig    abgebauten 

Nahrungsstoffe,  Ibid.,  77,  22-58. 
1907    Abderhalden,  Emil,  and  Casimir  Funk:  Beitrag  zur  Kenntniss  der  beim 

Kochen  von  Casein  mit    25%iger    Schwefelsaure    und    mit    starker 

Salzsaure  entstehenden  Spaltungsprodukte,  Ibid.,  53,  19-30. 

1906  Abderhalden,  Emil,  and  Andrew  Hunter:  Hydrolyse  des  im  Eigelb  des 

Huhnereies  enthaltenen  Proteins  ("Vitellin"),  Ibid.,  48,  505-512. 
1910    Abderhalden,  Emil,  and   Leo   Langstein:  Vergleichende   Untersuchung 

liber  die  Zusammensetzung  des  Caseins  aus  Frauen-  und  Kuhmilch, 

Ibid.,  66,  8-12. 
1909    Abderhalden,  Emil,  E.  S.  London,  and  Alfred  Schittenhelm :  Ueber  den 

Nuclienstoffwechsel  des  Hundes  bei  Ausschaltung  der  Leber  durch 

Anlegung  einer  Eckschen  Fistel,  Ibid.,  61,  413-418. 

1904  Abderhalden,  Emil,  and  Peter  Rona :  Futterungsversuche  mit  durch  Pan- 

kreatin,  durch  Pepsinsalzsaure  plus    Pankreatin    und    durch    Saure 
hydrolysierten  Casein,  Ibid.,  42,  528-539. 

1905  Abderhalden,  Emil,  and  Peter  Rona:  Ueber  die  Verwertung  der  Abbau- 

produkte  des  Caseins  in  tierischen  Organismus,  Ibid.,  44,  198-205. 

1906a  Abderhalden,  Emil,  and  Alfred  Schittenhelm:  Der  Ab-  und  Aufbau  der 
Nucleinsaure  im  thierischen  Organismus,  Ibid.,  47,  452-457. 

1906b  Abderhalden,  Emil,  and  Alfred  Schittenhelm:  Vergleichung  der  Zusam- 
mensetzung des  Caseins  aus  Frauen-,  Kuh-  und  Ziegenmilch,  Ibid.,  47, 
458-465. 

1902  Achard,  Ch.,  Ch.  Laubry,  and  L.  Thomas:  Contribution  a  l'etude  des 
phosphates  urinaires  dans  quelque  etats  aigus,  Bui.  et  mem.  Soc.  med. 
des  hopitaux  de  Paris,  3rd  series  19,  441-448. 


590  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1911  Achard,  H.  J.,  and  Harold  H.  Redneld:  On  the  Use  of  Lecithin  in 
Surgery,  New  York  Med.  Jour.  94,  779-782. 

1912a  Adler,  L. :  Ueber  organisch  und  anorganisch  gebundene  Phosphorsaure  im 
Bier  und  ihre  Beziehung  zu  Gerste  und  Malz  I,  Zeit.  ges.  Brauw.  35, 
181-184,  193-197,  210-213;  through  Zeit.  Unters.  d.  Nahrungs-  u. 
Genussmittel  24  (1912),  657. 

1912b  Adler,  L.:  Ueber  organisch  und  anorganisch  gebundene  Phosphorsaure  im 
Bier  und  ihre  Beziehung  zu  Gerste  und  Malz.  II.  Die  Phosphor- 
saureverhaltnisse  wahrend  der  Garung  und  in  der  Wiirze.  III.  Die 
Phosphorsaureverhaltnisse  in  Gerste,  Malz  und  Malzkeimen,  Ibid.,  35, 
246-250,  277-282,  293-298;  through  Zeit.  Unters.  d.  Nahrungs-  u. 
Genussmittel  25  (1913),  507. 

1909  Adler,  Max:  Ein  Fall  Phosphorsaure-Retention  und  deren  Beeinflussung 

durch  Medikamente,  Zentralbl.  ges.  Physiol,  u.  Path.  d.  Stoffwechs., 
N.  S.,  4,  242-248. 

1897  Adrian  and  Trillet:  Sur  le  phosphoglycerate  et  sur  la  dosage  des  phos- 

phoglycerates  neutres.  Jour,  de  pharm.  et  de  chim.  6,  481-485. 

1898  Adrian  and  Trillet:    Sur    les    phosphoglycerates    acides,    Compt.    rend. 

Acad,  des  sci.  126,  1215-1218. 
1872a  Aeby,  Carl:  Ueber  die  Constitution    des     phosphorsauren     Kalkes     der 

Knochen,  Jour.  f.  prakt.  Chem.,  113  (n.  s.  5),  308-311. 
1872b  Aeby,  Carl:  Ueber  die  nahren  Bestandtheile     des     Knochenphosphates, 

Ibid.,  114  (n.  s.  6),  169-171. 
1873a  Aeby,  Carl:  Ueber  die  Zusammensetzung  des  Knochenphosphates,  Cen- 

tralbl.  med.  Wissensch.,  11,  97-101. 
1873b  Aeby,  Carl:  Ueber  die  Beziehungen  des  Knochenknorpels  zum  Kalkphos- 

phate,  Ibid.,  11,  849-851. 
1873c  Aeby,  Carl:  Ueber  die  Metamorphose  der  Knochen,  Jour.  f.  prakt.  Chem., 

115  (n.  s.  7),  37-44. 
1874a  Aeby,  Carl:  Ueber  die  Konstitution  des  Knochenphosphates,  Chem.  Ber., 

7,  555-560. 
1874b  Aeby,  Carl:  Ueber  das  basische  Knochenphosphate,  Jour.  f.  prakt.  Chem. 

117  (n.  s.  9),  469-476. 
1905-6  Aeschbacher,  Siegfried :  Ueber  den    Einnuss  krankhaf ter    Zustande  auf 

den  Jod-   und  Phosphor  gehalt  der   normalen   Schilddruse,   Mitt,   aus 

Grenzgeb.  der  Med.  u.  Chir.,  15,  269-296. 
1893     Ajello,  G.,  and  A.  Solaro:  Le  ricambio  materiale  e  la  tossicata  dell'  urina 

nell'  inanizione  dell'  nomo,  La  Riforma  med.  IX,  Vol.  3,  No.  46-47,  p. 

542;  through  Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.,  24  (1894),  544,  545. 
1906     Albu,  Albert,  and  Carl  Neuberg:  Physiologie  und  pathologie  des  Miner- 

alstoffwechsels.      Berlin.  255  pages. 
1913     Alcindor,  J.:  Tuberculosis  and  Phosphorus  Metabolism,  Practioner  90, 

123-128. 

1910  Alcock,  N.  H.,  and  G.  R.  Lynch:  On  the  Relation  between  Physical, 

Chemical  and  Electrical  Properties  of  Nerves.  Part  III.  Total  Ash, 
Sulphates,  Phosphates,  Jour,  of  Physiol.  39,  402-410. 

1892  Alexander,  Carl:  Untersuchungen  uber  die  Nebennieren  und  ihre  Bezie- 
hungen zum  Nervensystem,  Beitrage  path.  Anat.  u.  allgem.  Path.,  11, 
145-189. 

1898  Alexander,  Franz:  Zur  Kenntniss  des  Caseins  und  seiner  peptischen 
Spaltungsprodukte,  Zeit.  physiol.  Chem.,  25,  411-429. 


PHOSPHORUS  METABOLISM  591 

1896  Alezais:  Note  sur  l'urine  normale  du  cobaye,  Compt.  rend.  Soc.  de  Biol., 

48,  213-214. 
1907     Allard,  E. :  Ueber  den  zeitlichen  Ablauf  der  Acidosekorper- Ausscheidung 

beim  Diabetes,  Arch.  exp.  Path.  u.  Pharm.,  57,  1-26. 
1907     Allers,  R.  A.,  and  S.  Bondi:  Ueber  das  Verhalten  des  Calciums  im  Blute 

bei  experimenteller  Saurevergiftung,  Biochem.  Zeit.,  6,  366-372. 
1906-6  Alquier,  J. :  Toxicite,  role  hygienique  et  therapeutique  des  sels  mineraux 

de  la  melasse  vis-a-vis  du  cheval,  Rep.  II  Congr.  internat.  de  l'aliment. 

yationelle  du  betail,  pp.  69-85. 
1904     Alsberg,   C.   L.:  Beitrage  zur  Kenntniss  der  Nucleinsaure,  Arch.   exp. 

Path.  u.  Pharm.,  51,  239-247. 
1889    Altmann,  Richard:  Ueber  Nucleinsauren,  Arch.  Anat.  u.  Physiol.,  physiol. 

Abt.,  pp.  524-536. 
1912    Altschul,  Julius:  Ueber  "Agfa-"Lecithin,  Biochem.  Zeit.,  44,  505,  506. 

1909  Alway,  F.  J.,  and  W.  L.  Hadlock:  The  Relation  of  the  Breaking  Strength 

of  Hog  Bones  to  the  Composition  of  the  Ash,  Neb.  Agr.  Exp.   Sta. 

Rept.  for  1909,  31,  32. 
1906    Amat,  Ch.:  La  demineralisation  phosphatee  de  l'organisme.     Cause  de 

1'  accroissement  de  la  tuberculose,  Bui.  gener.  de  therapeut.,  151,  411- 

414. 
1911a  Amberg,  Samuel,  and  Walter  Jones:  On  the  Application  of  the  Optical 

Method  to  a  Study  of  the  Enzymatic  Decomposition  of  Nucleic  Acids, 

Jour.  Biol.  Chem.,  10,  81-87. 
1911b  Amberg,  Samuel,  and  Walter  Jones :  Ueber  die    bei    der    Spaltung    der 

Nucleine  in  Betracht  kommenden  Fermente  mit  besonderer  Berticksich- 

tigung  der  Bildung  von  Hypoxanthin  in  der  Abwesenheit  von  Adenase, 

Zeit.  physiol.  Chem.,  73,  407-415. 

1897  Ames,  Delano,  and  A.  A.  Huntley:  The  Nature  of  the  Leucocytosis  Pro- 

duced by  Nucleinic  Acid:  A  Preliminary  Experimental  Study,  Jour. 
Amer.  Med.  Assoc,  29,  472-478. 

1910  Ames,  J.  W.:  The  Composition  of  Wheat,  Ohio  Agr.  Exp.  Sta.  Bui.  221. 

37  pp. 
1912    Ames,  J.  W.,  and  Geo.  E.  Boltz:  Nitrogen  and  Mineral  Constituents  of 

the  Alfalfa  Plant,  Ohio  Agr.  Exp.  Sta.  Bui.  247.    19  pp. 
1912    Ames,  J.  W.,  Geo.  E.  Boltz,  and  J.  A.  Stenius:  Effect  of  Fertilizers  on  the 

Physical  and  Chemical  Properties  of  Wheat,  Ohio  Agr.  Exp.  Sta.  Bui. 

243.      21  pp. 

1911  Anderson,  R.  J.:  Phytin  and  Phosphoric  Acid  Esters  of  Inosite,  N.  Y. 

Agr.  Exp.  Sta.,  Tech.  Bui.  No.  19;  also  Jour.  Biol.  Chem.,  11  (1912), 

471-488. 
1912a  Anderson,  R.  J.:  Phytin  and  Phosphoric  Acid  Esters  of  Inosite.  Second 

paper,  N.  Y.  Agr.  Exp.  Sta.,  Tech.  Bui.  No.  21;  also  Jour.  Biol.  Chem., 

12  (1912),  97-113. 
1912b  Anderson,  R.  J.:   The  Organic-Phosphoric   Acid   Compound   of  Wheat 

Bran,  N.  Y.  Agr.  Exp.  Sta.,  Tech.  Bui.  No.  22;  also  Jour.  Biol.  Chem., 

12  (1912),  447-464. 
1912c  Anderson,  R.  J. :  The  Organic-Phosphoric  Acid  of  Cottonseed  Meal,  N.  Y. 

Agr.  Exp.  Sta.,  Tech.  Bui.  No.  25;  also  Jour.  Biol.  Chem.,  13  (1912), 

311-323. 


592  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1914a  Anderson,  R.  J.:  Concerning  the  Organic  Phosphoric  Acid  of  Cottonseed 
Meal.    II.  (Fifth  papor  on  phytin)  Jour.  Biol.  Chem.  17,  141-150. 

1914b  Anderson,  R.  J.:  Concerning  Phytin  in  Oats.  (Sixth  paper  on  phytin) 
Ibid.  17,  151-163. 

1914c  Anderson,  R.  J.:  Concerning  Phytin  in  Corn.  (Seventh  paper  on  phytin) 
Ibid.  17,  165-170. 

1914d  Anderson,  R.  J. :  A  Contribution  to  the  Chemistry  of  Phytin.  I.  Compo- 
sition of  Barium  Phytate  and  Phytic  Acid.  II.  A  Study  of  the  Prop- 
erties of  Phytic  Acid  and  Its  Decomposition  Products  (Eighth  paper 
on  phytin)  Ibid.  17,  171-190. 

1898  Anger,  Albert:  Ueber  den  Umsatz  und  Ansatz  der  Aschenbestandtheile, 
vornehmlich  von  Kalk,  Magnesia,  Kali  und  Phosphorsaure,  bei  Milch- 
kiihen,  Inaug.  Diss.,  Heidelberg. 

1911  Anzilotti,  Giulio:  Leucociti  e  nucleinato  di  sodio,  Pathologica,  pp.  249- 
255;  through  Zentralbl.  Biochem.  u.  Biophys.,  12  (1911-12),  843. 

1908  Apelt,  F.,  and  O.  Schumm:  Untersuchungen  iiber  den  Phosphorsaurege- 
halt  der  Spinalflussigkeit  unter  pathologischen  Verhaltnissen,  Arch,  f . 
Psychiatrie  44,  845-855. 

1893  Apolant,  H.:  Ueber  die  Resorption  und  die  Apposition  von  Knochenge- 
webe  bei  der  Entwickelung  bosartiger  Knochentumoren,  Arch.  path. 
Anat.  u.  Physiol.,  131,  40-77. 

1903a  Araki,  T.:  Ueber  enzymatische  Zersetzung  -der  Nucleinsaure,  Zeit. 
physiol.  Chem.,  38,  84-97. 

1903b  Araki,  T.:  Ueber  die  Nucleinsaure  aus  der  Schleimhaut  des  Dunndarms, 
Ibid.,  38,  98-100. 

1908  Argiris,  Alfred:  Untersuchungen  iiber  Vogel-  und  Fischgehirne,  Ibid.,  57, 
288-295. 

1902  Aries,  Charles:  Contribution  a  l'etude  de  l'emploi  therapeutique  de  la 
lecithine  chez  les  vieillards,  These  de  Paris.      59  pp. 

1907  Arinkin,  M.:  Ueber  den  Einfluss '  einiger  anorganischer  und  organischer 

Sauren  auf  die  Autolyse  der  Leber,  Zeit.  physiol.  Chem.,  53,  192-214. 
1893     Arnstadt,  A.:  Das  Auftreten    der     Knochenkrankheit    beim     Rindvieh, 

Milch-Zeitung,  22,  510. 
1905     Aron,  Hans:  Ueber  den  Einfluss  der  Alkalien  auf  das  Knochenwachstum, 

Arch.  ges.  Physiol.,  106,  91,  92. 

1908  Aron,   Hans:   Kalkbedarf   und   Kalkaufnahme   beim    Saugling   und   die 

Bedeutung  des  Kalkes  fiir  die  Aetiologie  der  Rachitis,  Biochem.  Zeit. 
12,  28-77. 

1910a  Aron,  Hans:  Phosphorus  Starvation  with  Special  Reference  to  Beriberi 
I.,  Philippine  Jour,  of  Sci.,  B,  5,  81-97. 

1910b  Aron,  Hans:  Wachstum  und  Ernahrung,  Biochem.  Zeit.,  30,  207-226. 

1908  Aron,  H.,  and  K.  Frese:  Die  Verweitbarkeit  verschiedener  Formen  des 
Nahrungskalkes  zum  Ansatz  beim  wachsenden  Tier,  Biochem.  Zeit.,  9, 
185-207. 

1910  Aron,  Hans,  and  Felix  Hocson:  Phosphorus  Starvation  with  Special  Ref- 
erence to  Beriberi.  II.  Philippine  Jour,  of  Sci.,  B,  5,  98-122. 

1911a  Aron,  Hans,  and  Felix  Hocson :  Rice  as  Food :  Investigations  of  the  Nitro- 
gen and  Phosphorus  Metabolism  on  a  Diet  Consisting  Principally  of 
Rice  and  Other  Vegetable  Foodstuffs,  Ibid.,  6,  361-381. 


PHOSPHORUS  METABOLISM  593 

1911b  Aron,  Hans,  and  Felix  Hocson:  Reis  als  Nahrungsmittel.  Untersuch- 
ungen  iiber  den  N-  und  P2O5-  stoffwechsel  bei  Ernahrung  mit  Reis  und 
anderen  hauptsachlich  vegetabilischer  Nahrung,  Biochem.  Zeit.  32, 
189-203. 

1908  Aron,  Hans,  and  Robert  Sebauer:  Untersuchungen  iiber  die  Bedeutung 
der  Kalksalze  fur  den  wachsenden  Organismus,  Ibid.  8,  1-28. 

1891.  Arthus,  Maurice,  and  Calixte  Pages:  Recherches  sur  la  digestion 
gastrique  du  lait,  Mem.  Soc.  de  biol.,  43,  131-176;  through  Jahresb.  u. 
d.  Fortschr.  d.  Thierchem.  21,  139-141. 

1911  Aschenheim,  E.,  and  L.  Kaumheimer:  Ueber  den  Aschegehalt  der  Mus- 

kulatur  bei  Rachitischen,  Monatsschr.  Kinderheilk.,  10  (Originalien  2) 
435-438. 

1912  Aschner,  Bernhard:  Ueber    die    Funktion    der    Hypophyse,    Arch.  ges. 

Physiol.  146,  1-146.         19  pp.  titles  and  refs. 

1899  Ascoli,  Alberto:  Ueber  die  Plasminsaure,  Zeit.  physiol.  Chem.  28,  426-438. 
1900a  Ascoli,  Alberto:  Ueber  den  Phosphor  der  Nucleinstoffe,  Ibid.  31,  156-160. 
1900b  Ascoli,  Alberto:  Ueber  ein  neues  Spaltungsprodukt  des  Hefenucleins, 

Ibid.  31,  161-164. 

1900  Aulde,  John:  The  Clinical  Status  of  Nuclein,  Internat.  Med.   Gazette 

(N.  Y.),  Nov.,  1900;  through  Jour.  Amer.  Med.  Assoc.  35,2  1434. 
1907    Bachrach,  R.,  and  J.  Bartel:  The  Influence  of  Yeast  Nucleic  Acid  on  the 

Virulence  of  Tubercle  Bacilli,  Wien.    klin.    Wochenschr.    20,    1140; 

through  Chem.  Abs.  1,  2805. 
1900    Backhaus  and  W.  Cronheim:  Ueber  Zusammensetzung  der  Frauenmilch, 

Ber.  Landwirtsch.  Inst.  d.  Univ.  Konigsberg  5,  61-72. 
1907    Backus,  Lee  Seldon:  Experiments  with  Nuclein.       Abstracts  of  Work 

Done  in  the  Laboratory  of  Veterinary  Physiology  and  Pharmacology, 

No.  4.      N.  Y.  State  Vet.  Coll.,  Cornell  1907,  pp.  26-36. 

1881  Baginsky,  Adolf:  Ueber  den  Einfluss  der  Entziehung  des  Kalkes  in  der 

Nahrung  und  der  Fiitterung  mit  Milchsaure  auf    den    wachsenden 
Organismus,  Arch.  Anat.  u.  Physiol.,  physiol.  Abt.,  pp.  357,  358. 

1882  Baginsky,  Adolf:  Zur  Pathologie  der  Rachitis,  Arch.  path.   Anat.  u. 

Physiol.  87,  301-318. 

1883  Baginsky,  Adolf:  Ueber  die  phosphorsaureverbindungen  in    der    Milch, 

Zeit.  physiol.  Chem.  7,  354-361. 
1895     Baginsky,  A.,  and  Sommerfeld:  Zur  Chemie  der  kindlichen  Galle,  Arch. 

Anat.  u.  Physiol.,  physiol.  Abt,  p.  562. 
1910     Bahrdt,  Hans,  and  F.  Edelstein:  Das  Kalkangebot  in  der  Frauenmilch, 

Jahrb.  Kinderheilk.  72,  Erganzungsheft,  pp.  16-42. 
1912     Bain,  William:  Pharmacology  and  Therapeutics  of  Lecithin  and  Phytin, 

Lancet,  182,  918-921. 
1904     Bajmakov,  N.  F.:   (Organisierte  Eiweisskorper,  Eisen-  und  Phosphor- 

Gehalt  der  Kinder-  und  Kalbermuskeln  wahrend  verschiedener  Perioden 

der  ersten  Lebensjahre),  Diss.,  St.  Petersburg.     95  pp.         Through 

Biochem.  Centralbl.  2,  610. 
1887     Baldi,  Dario:  Einige  Beobachtungen  iiber  die  Verbreitung  des  Jecorins 

im   thierischen   Organismus,   Arch.   Anat.   u.   Physiol.,  physiol.   Abt., 

Suppl.,  pp.  100-108. 
1899-    Baldoni,  A.:  La  sostanze  grosse  ed  anorganische  della  tiroide,  Bui.  d. 
1900     reale  Accad.  med.  di  Roma  26,  218-224. 


594  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1896    Balke  and  Ide:  Quantitative?    Bestimmung     der     Phosphorfleischsaure, 

Zeit.  physiol.  Chem.  21,  380-386. 
1903    Balland,  M.:  Sur  les  quantites  de  phosphore  contenues  dans  les  farines, 

Compt.  rend.  Acad,  des  sci.  136,  332,  333. 
1907    Balland,  M.:  Sur  la  distribution  du- phosphore  dans  les  aliments,  Jour. 

de  pharm.  et  de  chim.,  ser.  6,  25,  9-12;  also  Bui.  Acad,  de  med.,  ser.  3, 

56  (1906),  612-614;  also  Compt.  rend.  Acad,  des  sci.  143  (1906),  969- 

970. 

1912  Ballerini,  G.:  Histochemische    Untersuchungen    iiber    Fettstoffe    und 

Lipoide  im  Plazentargewebe,  Arch.  Gynakol.  98,  156-186. 
1901a  Balthazard,  V.:  Les  lecithines  du  foie  a  l'etat  normal  et  pathologique, 

Compt.  rend.  Soc.  de  biol.  53,  922-924. 
1901b  Balthazard,  V.:  Les  lecithines  des  foies  gras  d'oie,  Ibid,  53,  1067,  1068. 

1913  Bamberg,  K.,  and  K.  Huldschinsky :  Ueber  angeborene  Knochenbriichig- 

keit,  Jahrbuch  f.  Kinderheilk.      78  SuppL,  214-266. 
1898    Bang,  Ivar:  Die  Guanylsaure  der  Pancreasdruse  und  deren  Spaltungs- 

produkte,  Zeit.  physiol.  Chem.  26,  133-159. 
1900a  Bang,  Ivar:  Bemerkungen  iiber  das  Nucleohiston,  Ibid.  30,  508-519. 
1900b  Bang,  Ivar:  Erwiderung,  Ibid.  31,  407-410. 
1900c  Bang,  Ivar:  Ueber  Parachymosin,  ein  neues    Labferment,    Arch.     ges. 

Physiol.  79,  425-441. 
1901a  Bang,  Ivar :  Chemische  und  physiologische  Studien  iiber  die  Guanylsaure, 

Zeit.  physiol.  Chem.  31,  411-427. 
1901b  Bang,  Ivar:  Chemische  und  physiologische  Studien  iiber  die  Guanylsaure. 

II.  Theil.    Physiologische  Studien,  Ibid.  32,  201-213. 

1903  Bang,  Ivar:  Chemische  Untersuchung  der  lymphatischen  Organe.    I,  II, 

III.  Beitrage  z.  chem.  Physiol,  u.  Path.  4,  115-138,  331-361,  362-  377. 

1904  Bang,  Ivar:  Chemische  Untersuchung  der  lymphatischen  Organe.     IV. 

Ibid.  5,  317-320. 

1907  Bang,  Ivar:  Biochemie  der  Zelllipoide.      I.      Ergebnisse  d.  Physiol.  6, 

131-186. 

1908  Bang,  Ivar:  Zur  charakteristik  der  Guanlysaure,     Beitrage     z.     chem. 

Physiol,  u.  Path.  11,  76-78. 

1909  Bang,  Ivar:  Biochemie  der  Zelllipoide.      II.      Ergebnisse  d.  Physiol.  8, 

463-523. 
1910a  Bang,  Ivar:  Ueber  die  Guanylsaure,  Zeit.  physiol.  Chem.  69,  167, 168. 
1910b  Bang,  Ivar:  Untersuchungen  iiber  die  Guanylsaure,  Biochem.  Zeit.  26, 

293-311. 
1911a  Bang,  Ivar:  Chemie  und  Biochemie  der  Lipoide,  Wiesbaden.    187  pp. 
(1911b)  Bang,  Ivar:  Ueber  die  chemische  Vorgang  bei  der  Milchgerinnung  durch 

Lab,  Skand.  Arch.  Physiol.  25  105-144;  through  Jahresb.  u.  d.  Fortsch. 

d.  Thierchem.  41  (1911),  221,  222. 
1912     Bang,  Ivar:  Erwiderungen.      I.      Ueber  "Agfa-"Lecithin,  Biochem.  Zeit. 

46,  500. 
1903     Bang,  Ivar,  and  C.  A.  Raaschon:  Ueber  die  Darstellung  der  Guanylsaure, 

Beitrage  z.  chem.  Physiol,  u.  Path.  4,  175-181. 

1905  Barbieri,  N.  Alberto:  Les  cerebrines  et  l'acide  cerebrique  preexistent  dans 

le  tissu  nerveaux  a  l'exclusion  du  protagon,  Compt.  rend.  Acad,  des 
sci.  140,  1551-1553. 
1909    Barbieri,  N.  Alberto:  Chemical  Composition  of  the  Nervous  System,  7th 
Internat.    Congr.  Appl.  Chem.,  Sect.  IV,  A2,  63-65. 


PHOSPHORUS  METABOLISM  695 

1910  Barbieri,  N.  Alberto:  Sur  la  non-existence  de  lecithines  libres  oil  com- 
bines dans  le  jaune  d'oeuf,  Compt.  rend.  Acad,  des  sci.  151,  405-407. 

1912  Barbieri,  N.  Alberto:  Sur  la  non-existence  des  lecithines  libres  ou  com- 
bines dans  le  jaune  d'oeuf  et  dans  les  structures  biologiques,  Ibid. 
155,  312-314. 

1900  Bardet,  G.:  Action  therapeutique  des  phosphoglycerates  acides,  Ibid. 
130,  956,  957. 

1905  Bardet,  M.  G. :  De  la  remineralisation  des  malades  en  etat  de  denutrition, 
Bui.  gener.  de  ther.,  med.,  chir.,  obstet.  et  pharm.  150,  106-111. 

1909a  Barille,  A.:  De  l'existence  des  carbonophosphates  dans  le  lait.  Leur 
precipitation  par  la  pasteurisation,  Compt.  rend.  Acad,  des  sci.  149, 
356-358. 

1909b  Barille,  A.:  De  l'existence  des  carbonophosphates  dans  le  lait;  leur 
precipitation  par  la  pasteurisation,  Jour,  de  pharm.  et  de  chim.,  ser.  6, 
30,  444-452. 

1910  Barille,  A.:   (Action  of  Dissociation  Products  of  Tricalcium  Carbono- 

phosphate  in  the  Genesis  of  Bony  Tissue  and  of  Various  Secretions 
Consisting  of  Phosphates  and  Carbonates  of  Calcium),  Ibid.,  ser.  7,  1, 
342-348;  377-380;  through  Chem.  Abs.  5  (1911),  514. 

1911  Barille,  A. :  Formation  du  tartre  dentaire  par  les  produits  de  dissociation 

des  carbonophosphates  de  la  salive,  Ibid.,  ser.  7,  3,  582-585. 
1904    Barnstein,  F.:  Ueber  Futterknochenmehl  und  dessen  Verfiitterung,  Deut. 

landwirtsch.  Presse  31,  771,  772. 
1899     Barratt,  Wakelin:  On  the  Amount  of  Water  and  Phosphorus  Contained 

in  the  Cerebral  Hemispheres  and  Spinal  Cord  in  General  Paralysis  of 

the  Insane  and  in  Other  Conditions,  Archives  of  Neurology  1,  207-222. 
1898    Basch,  Earl:  Die  Entstehung  des  Caseins  in  der  Milchdruse,  Jahrb.  Kin- 

derheilk.,  N.  F.  47,  90-104. 
1903     Basch,  Karl:  Die  Physiologie  der     Milchabsonderung,     Ergebnisse     d. 

Physiol.  2,  1st  Abt.,  326-376. 
1908     Baskoff,  A.:  Ueber  das  Jecorin  und  and  ere  lecithinartige  Produkte  der 

Pferdeleber,  Zeit.  physiol.  Chem.  57,  395-460. 
1909a  Baskoff,  A.:    Ueber  Lecithinglykose    in    Vergleich    zum    Jecorin    der 

Pferdeleber,  Ibid.  61,  426-453. 
1909b  Baskoff,  A.:  Ueber  Lecithin  und  Jecorin  der  Leber  normaler  und  mit 

Alcohol  vergifteter  Hunde,  Ibid.  62,  162-172.  . 
1858a  Baud,  Victor:  Emploi  therapeutique  de  composes  phosphores  extraits  de 

la  moelle  allongee  des  mammiferes  herbivores,  Compt.  rend.  Acad. 

des  sci.  46,  858,  859. 
1858b  Baud,  Victor.:  Nouvelles  etudes  sur  les  corps  gras  phosphores  extraits 

de  la  moelle  allongee  des  mammiferes  herbivores,  faites,  d'apres  les 

indications  de  M.  Mege-Mouries.      Paris  51  pp. 
1910    Baudran,  G.:  Bacilles  de  Koch.      Milieux  aux  glycerophosphates.  Doses 

maxima  de  fer  et  de  manganese,  Compt.  rend.  Acad,  des  sci.  150,  1200- 

1202. 
1907    Bauer,  Friedrich:  Die  Konstitution  der  Inosinsaure  und  die  Muskelpen- 

tose,  Beitrage  z.  chem.  Physiol,  u.  Path.  10,  345-357. 
1868    Bauer,  H.:  Knochenerweichung  beim  Rinde,  Wochenschr.  f.  Thierheilk. 

U.  Viehzucht  12,  334,  335. 


596  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

(1911)  Bauer,  J.:  Ueber  den  Befund  von  Kuhmilchkase'in  in  Sauglingssttihlen, 
Monatsschr.  f.  Kinderheilk.  10,  239-243;  through  Jahresb.  u.  d. 
Fortschr.  d.  Thierchem.  41  (1911),  298. 
1911  Bauer,  J.,  and  St.  Engel:  Ueber  die  chemische  und  biologische  Differ- 
enzierung  der  drei  Eiweisskorper  in  der  Kuh-  und  Frauenmilch, 
Biochem.  Zeit.  31,  46-64. 
1871    Bauer,  Jos.:  Der  Stoffumsatz  bei  der  Phosphorvergiftung,  Zeit.  Biol.  7, 

61-85. 
1878     Bauer,  Jos.:  Ueber  die  Eiweisszersetzung  bei  Phosphorvergiftung,  Ibid. 

14,  527-541. 
1911     Bauer,  T.:  Ueber  das  Verhalten  der  Epithelkorperchen  bei  der  Osteo- 
malacic, Frankfurter  Zeit.  Path.  7,  231-237. 
1913     Baumann,  A.:  Ueber  den  stickstoffhaltigen  Bestandteil  des  Kephalins, 

Biochem.  Zeit.  54,  30-39. 
1885    Baumstark,  F.:  Ueber  eine  neue  Methode,     das     Gehirn    chemisch    zu 
erforschen,  und  deren  bisherige  Ergebnisse,  Zeit.  physiol.   Chem.   9, 
145-210. 
1904     Bayliss,  W.  M. :  The  Kinetics  of  Tryptic  Action,  Archives  des  sci.  biol.  11, 

Suppl.,  261-296.       St.  Petersburg. 
1913     Bayliss,  W.  M.:  Researches  on  the  Nature  of  Enzyme  Action.      III.  The 

Synthetic  Action  of  Enzymes,  Jour,  of  Physiol.  46,  236-266. 
1903     Bayon,  P.  G.:  Erneute  Versuche  iiber  den  Einfiuss   des   Schilddrusen- 
verlustes  und  der  Schilddrusenfiitterung  auf  die  Heilung  von  Knochen- 
bruchen,  Verhandl.  Physik.-Med.  Gesellsch.   zu  Wiirzburg,  N.  F.   35, 
249-311. 
1893     Bechamp,  A.:  Sur  la  caseine  et  le  phosphore  organique  de  la  caseine, 
Compt.  rend.  Acad,  des  sci.  117,  1085-1088;  also  Bui.  Soc.  chim.  de 
Paris  61  (ser.  3,  Vol.  11)  (1894),  152-176. 
1901     Bechhold,  Heinrich:  Ueber  Phosphorsaureester    von    Eieralbumin,    Zeit. 

physiol.   Chem.  54,  122-127. 
1844     Becquerel,  Louis  Alfred,  and  A.  Rodier:  Recherches  sur  la  composition 
du  sang  dans  l'etat  de  sante  et  dans  l'etat  de  maladie,  Paris.      127  pp. 
Trans,  by  Eisemann  1845,  162  pp. 
1907    Benedict,  Francis  Gano:  The  Influence  of  Inanition  on  Metabolism,  Car- 
negie Inst.  Pub.  77. 
1870     Beneke,  F.  W.:  Zur  Wiirdigung  des  Phosphors.  Kalkes  in  physiolog.  und 
therapeut.  Beziehung,  Marburg.      48  pp.      See  Jahresb.  d.  in-  u.  aus- 
landisch.  ges.  Med.  151  (1871),  137-139. 
1906     Bensley,  R.  R.:  An  Examination  of  the  Methods  for  the  Microchemical 
Detection  of  Phosphorus  Compounds  Other  than  Phosphates  in  the 
Tissues  of  Animals  and  Plants,  Biological  Bui.  10,  49-65. 
1910    Benson,  C.  C,  M.  B.  Tamblyn  and  Students:  A  Day's  Metabolism,  Jour. 

Home  Economics  2,  658. 
1890     Beraz,  Heinrich:  Ueber  die  Bedeutung  des  Kalkes  fur  die  Zahne,  Zeit. 

Biol.  27,  386-397. 
1910a  Berg,  Ragnar:  Ueber  das  Ausscheidung  von  per  os  eingeftihrten  Phos- 

phaten,  besonders  der  Calciumphosphate,  Biochem.  Zeit.  30,  107-142. 
1910b  Berg,  Ragnar:  Ueber  Phosphorsaurestoffwechsel,  Chem.  Zeitung  34,  1049. 
1911a  Berg,   Ragnar:   Eine   Fehlerquelle  bei  Versuchen  iiber  Phosphorsaure- 
stoffwechsel, Zeit.  physikal.  u.  diatet.  Ther.  15,  98-100. 


PHOSPHORUS  METABOLISM  597 

1911b  Berg,  Ragnar:  Schlusswort,  Ibid.  15,  240-242. 

1910    Berg,  R.,  and  C.  Rose:  Ueber  den  Mechanismus  der  Beeinflussung  der 

korperlichen  Entwicklung  durch  die  Wasserharte,  Biochem.  Zeit.  27, 

204-222. 
1906-7  Berg,  William  N.,  and  William  J.  Gies:  Studies  of  the  Effects  of  Ions 

on  Catalysis,  with  Particular  Reference  to  Peptolysis  and  Tryptolysis, 

Jour.  Biol.  Chem.  2,  489-546. 
1898a  Bergell,  Peter:  Ueber  den  Phosphorsaurestoffwechsel  unter  normalen  und 

pathologischen  Verhaltnissen,  Fortschritte  der  Med.  16,  1-18. 
1898b  Bergell,  Peter:  Die  Bedeutung  der  Phosphorsaure  im  menschlichen  und 

thierischen  Organismus,  Inaug.  Diss.,  Berlin. 

1900  Bergell,  Peter:  Darstellung  des  Lecithins,  Ber.  deut.  chem.  Gesell.  33,  II, 

2584-2586. 

1901  Bergell,  Peter:  Ueber  das  Spaltung  des  Lecithins  durch  den  bei  voll- 

standigem  Darmverschluss  abgesonderten  Darmsaft,  Centralbl.  allgem. 

Path.  u.  path.  Anat.  12,  633,  634. 
1905    Bergell,  Peter,  and  A.  Braunstein:  Ueber  das  Lecithin  und  Bromlecithin, 

Ther.  der  Gegenwart  46  (N.  S.,  7),  156-160. 
1908    Bergell,  Peter,  and  Leo  Langstein:  Beitrage  zur  Chemie  der  Frauen- 

milch.    I.  Mitt.  Unterschiede  zwischen  dem  Kasein  der  Frauen-  und 

Kuhmilch,  Jahrb.  Kinderheilk.  68,  568-576. 

1901  Bergmann,  W.:  Ueber  die  Ausscheidung  der  Phosphorsaure  bei  Fleisch- 

und  Pflanzenfressern,  Arch.  exp.  Path.  u.  Pharm.  47,  77-81. 
1908    Berkley,  H.  J. :  Action  of  Lecithin  in  Exophthalmic  Goiter,  Johns  Hopkins 

Hosp.  Bui.,  Sept.,  1908;  through  Jour.  Amer.  Med,  Assoc.  51,  1464. 
1903a  Bernard,  Leon,  Bigart  and  H.  Labbe:  Sur  la  secretion  de  lecithine  dans 

les  capsules  surrenales,  Compt.  rend.  Soc.  de  biol.  55,  120-122. 
1903b  Bernard,  Leon,  Bigart  and  H.  Labbe:  Importance  de  la  lecithine  dans  les 

fonctions  de  la  capsule  surrenale  et  sa  secretion  par  cet  organe,  La 

presse  med.  11,  119,  120. 
1912    Bernardini,  Luigi:  (Chemical  Composition  of  the  Rice  Embryo),  Atti. 

accad.  Lincei  21,  I,  283-289;  through  Chem.  Abs.  6  (1912),  1171. 
1908    Bernazky,  Stanislaw  Stanislawovitch :  (Das  Lezithin  des  Knochenmarks 

normaler  und  immunisierter  tiere  und  die  Verteilung  des  Phosphats  im 

Organismus),  St.  Petersburg.      60  pp.;  through  Biochem.  Centralb.  7 

(1908),  827. 

1898  Bernstein:  Die    Oophorinbehandlung    bei    Osteomalacic,    Munch,    med. 

Wochenschr.  45, 1,  427,  428. 
1878    Bertram,  Julius:  Ueber  die  Ausscheidung  der  Phosphorsaure  bei  den 
Pflanzenfressern,  Zeit.  Biol.  14,  335-382. 

1899  Bertz,  F. :  Ueber  die  chemische  Zusammensetzung  der  Zahne,  Inaug.  Diss., 

Wiirzburg.    36  pp.;  through  Jahresb.  u.  d.  Fortschr.  d.  Thierchem.  30 
(1900),  457,  458. 
1816    Berzelius,  Jacob:  Untersuchungen  uber  die  Zusammensetzung  der  Phos- 
phorsaure, der  phosphorigen  Saure  und  ihrer  Salze,  Gilbert's  Annalen 
der  Physik  53  (new  ser.  23),  393-446. 

1902  Bethe,  Albrecht:  Ueber  einige  Educte  des  Pferdegehirns,  Arch.  exp.  Path. 

u.  Pharm.  48,  73-86. 
1858    von  Bezold,  A. :  Das  chemische  Skelett  der  Wirbelthiere,  Zeit.  wissensch. 
Zool.  9,  240-269;  through  Jahresb.  ii.  d.  Fortschr.  d.  Chem.  u.  verwand- 
ter  Theile  andere  Wissensch.  1858,  553,  554. 


598  OHIO  EXPEEIMENT  STATION:  TECHNICAL  BUL.  5 

1844     von  Bibra,   Ernst:   Chemische  Untersuchungen  liber   die  Knochen  und 

Zahne  des  Mensehen  und  der  Wirbelthiere,  mit  Rucksichtnahme  auf 

ihre  physiologischen  und     pathologischen     Verhaltnisse,  Schweinfurt. 

430  pp.     Through  Schrodt,  1876. 
1853     von  Bibra,  Ernst:  Ueber  das  Gehirn,  Annalen  der  Chem.  u.  Pharm.  57, 

201-224. 
1854a  von  Bibra,  Ernst:  Vergleichende  "Untersuchungen  iiber  das  Gehirn  des 

Mensehen  und  der  Wirbelthiere,  Mannheim.  134  pp. 
1854b  Von  Bibra,  Ernst:  Ueber  das  Ruckenmark  und  die  Nerven/ Annalen  der 

Chem.  u.  Pharm.  91,  1-33. 
1911a  Bickel,  A.:  Zur  Kenntnis  des  Lezithinstoffwechsels,  Internat.  Beitrage  z. 

Path.  u.  Ther.  der  Ernahrungsstorungen  3,  171-179. 
(1911b)  Bickel,  A.:  Ueber  Mastkuren,  Med.  Klinik  (Berlin)  12,  441,442;  through 

Jahresb.  u.  d.  Fortschr.  d.  Thierchem.  41  (1911),  473. 
1852     Bidder,  F.,  and  C.  Schmidt:  Die  Verdauungssafte  und  der  Stoffwechsel. 

Eine  physiologisch-chemische  Untersuchung,  Mitau   u.  Leipzig.  413  pp. 
1874     Biedert,   Philipp:   Neue   Untersuchungen   und  klinische   Beobachtungen 

iiber  Mensehen-  und  Kuhmilch  als  Kindernahrungsmittel,  Arch.  path. 

Anat.  u.  Physiol.  60,  352-379. 
1880     Biedert,    Philipp:    Die   Kinderernahrung   im    Sauglingsalter,    Stuttgart. 

372  pp.  153  refs.      Later  editions;  1893,  1897. 
1884     Biedert,  Philipp:  Untersuchungen  iiber  die  chemischen  Unterschiede  der 

Mensehen-  und  Kuhmilch,  Stuttgart.  69  pp. 
1887     Biedert,  Philipp:  Ueber  die  Eiweisskorper  der  Mensehen-  und  Kuhmilch, 

Deut.  med.  Wochenschr.  13,  105,  106;  see  also  Schroter,  1887. 

1897  Biedert,  Philipp:  Die  Kinderernahrung  im  Sauglingsalter  und  die  Pflege 

von  Mutter  und  Kind,  Stuttgart. 
1912     Bienenfeld,  Bianca:  Beitrag  zur  Kenntnis  des  Lipoidgehaltes  der  Pla- 
centa, Biochem.  Zeit.  43,  245-255. 

1909  Biernacki,  E.:  (Ueberernahrung     und     Mineralstoffwechsel),     Tygodnik 

lekarski  4,  197-206  (Polish);  through  Zentralbl.  ges.  Physiol,  u.  Path.  d. 
Stoffwechsels  10  (new  ser.  4)  (1909),  449-455;  481-496;  and  11  (new 
ser.  5)   (1910),  240. 

1910  Biernacki,  E.:  Ueber  die  Beziehungen  zwischen  dem  Umsatz  mineralischer 

und  organischer  Nahrungsstoffe,  Wien.  klin.  Wochenschr.  23,  850-854. 

1898  Biffi,  Ugo:  Zur  Kenntniss  der  Spaltungsprodukte  des  Caseins  bei  der 

Pankreasverdauung,  Arch.  path.  Anat.  u.  Physiol.  152,  130-157. 

1904  von  Bilgorajski,  Marian:  Klinische  Erfahrungen  mit  Protylin  und  dessen 
Eisen-  und  Bromkombinationen,  Wien.  klin.  Rundschau  18,  188-190; 
206-208. 

1903a  Billon,  F.,  and  H.  Stassano:  Sur  la  maniere  d'etudier  l'action  des  com- 
poses phosphores  organique  naturels  et  synthetiques,  Comp.  rend.  Soc. 
de  biol.  55,  276. 

1903b  Billon,  F.,  and  H.  Stassano:  Action  de  quel  que  composes  phosphores  sur 
la  nutrition,  Ibid.  55,  277-279. 

1899  Bing,  H.  J.:  Untersuchungen  iiber     die     reducirenden     Substanzen     im 

Blute,  Skand.  Arch.  Physiol.  9,  336-411. 
1901     Bing,  H.  J.:  Ueber  Lecithinverbindungen,  Ibid.  11,  166-175. 
1910     Bircher,    E.:    Zur   Wirkung    der   Thyreoidintabletten   auf   das   normale 

Knochenwachsthum,  Arch.  klin.  Chir.  91,  554-566. 


PHOSPHORUS  METABOLISM  599 

1909    Birk,  Walter:  Untersuchungen  iiber  den  Einfluss  des  Phosphorlebertrans 

auf  den  Mineralstoffwechsel  gesunder    und    rachitischer     Sauglinge, 

Monatsschr.  Kinderheilk.  7,  450-475. 
19H     Birk,  Walter:  Zur  Physiologie  des  neugeborenen  Kindes.    IV.  Mitteilung. 

Der  Stoffwechsel  des     Kindes  wahrend     der  ersten     Lebenstage  bei 

kiinstlicher  Ernahrung,  Ibid.  10,  1-11. 
1867     Bischoff,  Ernst:  Ueber  die  Ausscheidung  der  Phosphorsaure  durch  den 

ThierkBrper,  Zeit.  Biol.  3,  309-323. 
1894     von  Bitto,  Bela:    Ueber    die     Bestimmung     des     Lecithingehaltes     der 

Pflanzensamen,  Zeit.  physiol.  Chem.  19,  488-498. 
1913     de  la  Blanchardiere,  P.:  Ueber  die  Wirkung  der  Nuclease,  Ibid.  87,  291- 

309. 
1897a  Blauberg,  Magnus:  Ueber  die  chemische     Zusammensetzung     einiger 

"Nahrsalz,"  nebst  kurzen    Bemerkungen    iiber    die    Bedeutung    der 

Mineralstoffe  fiir  den  Organismus,  Arch.  Hygiene  30,  95-124. 
1897b  Blauberg,  Magnus:  Weitere  Untersuchungen  iiber  Kindernahrungsmittel, 

nebst  kurzen  Bemerkungen  iiber  die  mikroscopische  und  bacteriolog- 

ische  Prufung  derselben,  Ibid.  30,  125-155. 
1897c  Blauberg,  Magnus:  Ueber  die  Mineralbestandtheile  der  Sauglingsfaces 

bei  natiirlicher  und  kiinstlicher  Ernahrung  wahrend  der  ersten  Lebens- 

woche,  Ibid.   31,   115-141;   also   Inaug.   Diss.,   Berlin,   1897,   abstract 

Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  27  (1897),  382,  383. 
1900a  Blauberg,  Magnus:  Experimented  Beitrage  zur  Frage  iiber  den  Mineral- 
stoffwechsel bei  kunstlich  ernahrten  Saugling,  Zeit.  Biol.  40   (N.  S. 

22),  1-35. 
1900b  Blauberg,  Magnus:    Ueber    den    Mineralstoffwechsel    beim    naturlich 

ernahrten  Saugling,  Ibid.  40  (N.  S.  22),  36-53. 
1906    Bloch,  Bruno:  Die  Umwanglung  der  Purinkorper  im  Saugetierorgan- 

ismus,  Biochem.  Centralb.  5,  521,  561,  817,  873. 
1900    Bloch,  Ernst:  Ueber  das  Plasmon   (Caseon)   als  Eiweissersatz,     nebst 

Beitragen  zur  Lehre  vom  Eiweissstoffwechsel,  Zeit.  diatet.  u.  physikal. 

Ther.  3,  482-504. 
1861     Blondlot:  Sur  la  recherche  toxicologique  du  phosphore  par  la  coloration 

de  la  flame,  Compt.  rend.  Acad,  des  sci.  52,  1197-1200. 
1912-    Bliihdorn:  Eine  Demonstration  des  Einflusses  der    Reaktion    auf    den 
1913      Umsatz  von  Kalk  und  Phosphorsaure  im  Dickdarm  der  Sauglings, 

Monatsschr.  Kinderheilk.,  Originalen,  11,  68-79. 
1896    Blumenthal,  Ferd.:  Ueber  die  Produkte  der  bakterischen  Zersetzung  der 

Milch,  Arch.  path.  Anat.  u.  Physiol.  146,  65-85. 
1879    Blyth,  A.  Winter:  The  Composition  of  Cow's  Milk  in  Health  and  Disease, 

Jour,  of  the  Chem.  Soc.  35,  530-539. 
1896    Boddaert,  A.:  Contribution  a  l'etude  de  Taction  des  hypophosphites  sur 

la  nutrition,  Archives  de  pharmacodynamic  2,  195-207. 
1849    Bocker,  Friedrich  Wilhelm:  Beitrage  zur  Heilkunde,  insbesondere  zur 

Krankheits-,  Genussmittel-  und  Arzneiwirkungs-Lehre,  nach  eigenen 

Untersuchungen,  vol.  I,  139-187. 
1911     Bogdanow,  B.  A.:  (Zur  Frage  iiber  die  Ausscheidung  des  organischen 

Phosphors  (im  Ham)     bei    einigen    Infektionskrankheiten),    Russki 
Wratsch  1911,  Nr.  17,  18,  19,  20;  through  Jahresb.  U.  d.  Fortschr.  d. 

Thierchem.  41  (1911),  485. 


600  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1877    Bokay,  A.:  Ueber  die  Verdaulichkeit  des  Nucleins  und  Lecithins,  Zeit. 

physiol.  Chem.  1,  157-164. 
1910a  Bolle,  Alwin:  Ueber  den  Lecithingehalt  des  Knochenmarks  von  Mensch 

und  Haustieren,  Inaug.  Diss.,  Erlangen.  25  pp. 
1910b  Bolle,  Alwin:  Ueber  den  Lecithingehalt  des  Knochenmarks  von  Mensch 

und  Haustieren,  Biochem.  Zeit.  24,  179-190. 

1902  Bonanni,  A.:   (Die  Phosphorfleischsaure  der  Muskeln  bei  Veratrinver- 

giftung),  Accad.  med.  di  Roma  29,  through  Jahresb.  u.  d.  Fortschr.  d. 

Thierchem.  32  (1902),  537,  538. 
1906    Bonanni,  A.:  Ein  weiterer  Beitrag  zur  Kenntnis  der  menschlichen  Galle. 

Die  Gegenwart  gebundener  Glykuronsaure  in  der  Galle;  Ausscheidung 

der  Borneol-Glykuronsaure  durch  dieselbe,  Bullett.  d.  reale  Accad.  med. 

di  Roma.  Anno.  32;  through  Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  36 

(1906),  468-470. 
1894a  Bongartz:  Ueber  seuchenartiges  Auftreten  der  Knochenbriichigkeit  im 

Fruhjahr  und  Sommer  dieses  Jahres,  Berlin,  tierarzte  Wochenschr. 

435-437. 
1894b  Bongartz:  Ueber  seuchenartiges  Auftreten  der  Knochenbriichigkeit  im 

Fruhjahr  und  Sommer  dieses  Jahres,  Fiihling's  Landwirtsch.  Zeitung 

43,  666-668;  see  Grandeau,  1905. 
1913     Bonnamour,  Albert  Badolle,  and    Escallon:    Decalcification    et    lesions 

osseuses  chez  le  lapin  sous  l'infiuence  du  lactose  en  injections  intra- 

veineuses,  Compt.  rend.  Soc.  de  biol.  74,  1106,  1107. 

1903  Bonnetat,  L.:  Action  des  engrais  phosphates  sur  la  composition  chem- 

ique  des  fourrages,  Jour,  d'agri.  prat.  5,  343-345. 
1906    Boos,  William  F.:  Ueber  Darstellung  und  Zusammensetzung  der  Myko- 

nukleinsaure  aus  Hefe,  Arch.  exp.  Path.  u.  Pharm.  55,  16-20. 
1909     Boos,  William  F.:  On  the  Reducing  Compound  of  Yeast  Nucleic  Acid, 

Jour.  Biol.  Chem.  5,  469-475. 
1838     Bopp:  Stimmen  liber  die  als  Knochenbriichigkeit  erkannte  Krankheit  des 

Rindviehes  in  der  Provinz  Rheinheffen,  Magazin.  f.  d.  ges.  Thierheilk. 

4,  322-350. 
1902a  Bordas,  F.,  and  de  Raczkowski:  Variation  de  l'acide  phosphorique  suivant 

l'age  du  lait,  Compt.  rend.  Acad,  des  sci.  135,  302,  303;  also  Annales 

de  chim.  anal.  7  (1902),  370,  371. 
1902b  Bordas,  F.,  and  de  Raczkowski :  De  l'infiuence  de  l'ecremage  sur  la  repar- 
tition des  principaux  elements  constitutifs  du  lait,  Compt.  rend.  Acad. 

des  sci.  135,  354,  355:  also  Annales  de  chim.  anal.  7  (1902),  372,  373. 
1903     Bordas,  F.,  and  de  Raczkowski:  Diminution  du  taux  des  lecithines  dans 

les  laits  chauffes,  Compt.  rend.  Acad,  des  sci.  136,  56,  57;  also  Annales 

de  chim.  anal.  8  (1903),  168,  169. 
1909     Bornstein,  Arthur:  Die  chemische  Zusammensetzung    des     Blutes    bei 

progressiver  Paralyse,  Monatsschr.  Psychiat.  u.  Neurol.  25,  160-168. 
1911     Bornstein,  Arthur :  Ueber  die  Lecithinamie  der  Geisteskranken,  Zeit.  ges. 

Neurol,  u.  Psychiat.,  Originalien  6,  605-608. 

1905  Bornstein,  Karl:  Ueber  den  Schwefel-     und     Phosphorstoffwechsel     bei 

abundanter  Eiweisskost,  Arch.  ges.  Physiol.  106,  66-79. 

1906  Borri,  Andrea :  Untersuchung  iiber  die  phosphorhaltigen  Bestandteile  der 

Darmschleimhaut,  Arch.  Verdauungskrankheiten  12,  301-307;  also 
Studio  sui  composti  fosforati  della  mucosa  intestinale,  La  Clinica  med. 
ital.,  Milano  45,  458-464. 


PHOSPHORUS  METABOLISM  601 

1910  Borrino,  Angiola:  Sulla  nucleasi  della  ghiandola  mammaria,  Archivio 

di  fisiologia,  8,  73-80. 

1911  Borrino,  Angiola:  Sull'  origine  della  caseina  del  latte,  Rivista  di  clinica 

pediatrica   9,   211-220;    through   Zentralbl.    Biochem.   u.    Biophys.    13 

(1912),  744,  745.      44  refs. 
1911     Borschim,  S.:  Ueber  den  Einfluss  des  Lecithins  auf  die  Resorption  der 

Haut,  Biochem.  Zeit.  35,  471-477. 
1907     Bosworth,  Alfred  W.:  Chemical   Studies  of  Camembert  Cheese,  N.  Y. 

Agr.  Exp.  Sta.,  Tech.  Bui.  No.  5.      Pp.  23-39. 
1913     Bosworth,  Alfred  W. :  The  Action  of  Rennin  on  Casein,  Jour.  Biol.  Chem. 

15,  231-236;  also  N.  Y.  Agr.  Exp.  Sta.  (Geneva),  Tech.  Bui.  No.  31, 

7  pp. 
1913     Bosworth,  Alfred  W.,  and  Lucius  L.  Van  Slyke:  Preparation  and  Com- 
position of  Basic  Calcium  Caseinate  and  Paracaseinate,  Jour.  Biol. 

Chem.  14,  207-210. 
1903     Bottazzi,  F.:  Proprieta  di  nucleoproteidi  estratti  dalla  placenta  muliebre 

(Properties  of  Nucleoprotein     Extracted     from     Human     Placenta.), 

Bollett.  d.  r.  Accad.  med.  di  Genova  18,  247-250. 
1846a  Boussingault,  M.:  Sur  le  developpement  de  la  substance  minerale  dans 

le  systeme  osseux  du  pore,  Annales  de  chim.  et  de  phys.,  3rd  ser.,16, 

486-493. 
1846b  Boussingault,  M.:  Untersuchungen  Tiber  die  Entwickelung  der  mineral- 

ischen  Substanzen  in  dem  Knochensystems  des  Schweins,  Annalen  der 

Chem.  u.  Pharm.  59,  322-330. 
1830     Braconnot,  H.:  Memoire  sur  le  caseum  et  sur  le  lait;  nouvelles  ressources 

qu'ils  peuvent  offrir  a  la  societe,  Annales  de  chim.  et  de  phys.,  2nd 

ser.,  43,  337-351. 
1907     Braddon,  W.  L.:  The  Cause  and  Prevention  of  Beri-Beri,  London  and 

New  York.      XIII  +  544  pp.      Through  Exp.  Sta.  Record  24  (1911), 

66,  67. 
1913    Brahm,  C:  Nukleinsauren  und  Spaltprodukte;  Oppenheimer's  Handbuch 

der  Biochemie,  Erganzungsband,  80-104. 
1902     Brand,  J.:   Beitrag  zur  Kenntniss  der  menschlichen  Galle,  Arch.   ges. 

Physiol.  90,  491-522.      89  refs. 
1903-4  Braunstein,  A. :  Beobachtungen  uber  die  Ausscheidung  der  Chloride,  der 

Phosphorsaure,  des  Stickstoffs  und  Ammoniaks  bei  Carcinom,  Zeit.  f. 

Krebsforschung  1,  199-224. 

1911  Breaudat,  L.,  and  Denier:  The  Use  of  Rice  Bran  in  the  Prevention  and 

Cure  of  Beriberi,  Annales  de  lTnst.  Pasteur  25,  167-189. 

1851a  Breed,  D.:  Ueber  den  Gehalt  des  normalen  Urins  an  Phosphorsaure, 
Annalen  der  Chem.  u.  Pharm.  78  (n.  s.  2),  150-157. 

1851b  Breed,  D.:  Analyse  der  Asche  des  menschlichen  Gehirns,  Ibid.  80,  124. 

1891  Breisacher,  Leo:  Zur  Physiologie  des  Schlafes,  Dubois-Reymond's  Arch. 
Physiol.  321-334. 

(1911)  Brennemann,  Joseph:  (Ein  Beitrag  zur  Kenntniss  der  Ursache  und 
Herkunft  der  harten  Brockel  im  Sauglingsstuhl),  Amer.  Jour.  Dis- 
eases of  Children  1,  341-359;  through  Jahresb.  u.  d.  Fortschr.  d.  Thier- 
chem.  41  (1911),  298. 

1912  Brossa,  G.  A.:  Ueber  die  biologische  Wertigkeit  der  a-Nucleinsaure, 

Arch.  Anat.  u.  Physiol,,  physiol.  Abt.,  191-196. 


602  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1892a  Brown-Sequard:  Effets  physiologique  d'un  liquide  extrait  des  glandes 

sexuelles  et  surtout  des  testicules,  Compt.  rend.  Acad,  des  sci.  114, 

1237. 
1892b  Brown-Sequard:   Effets  produits   sur   de  nombreux  morbides  par  des 

injections  sous-cutanees  d'un  extrait  liquide  retire  des  testicules,  Ibid. 

114,  1318,  116,  856. 
1890     Brubacher,  Heinrich:  Ueber    den    Gehalt    an    anorganischen    Stoffen, 

besonders  an  Kalk,  in  den  Knochen  und  Organen  normaler  und  rachit- 

ischer  Kinder,  Zeit.  Biol.  27,  517-549. 
1908    Bruck,  A.  W.:  Ueber  den  Mineralstoffwechsel  beim  kiinstlich  genahrten 

Saugling,  Monatsschr.  Kinderheilk.    6,  570-579. 

1908  Brugsch,   Theodor:   Zur   Physiologie  und   Pathologie   des   Nukleinstoff- 

wechsels,  Med.  Klinik,  Berlin,  4,  II,  1841-1843. 

1909  Brugsch,  Theodor:  Zur  Stoffwechselpathologie  der  Gicht.  VIII.  Mitteil- 

ung.  Die  Harnsaure  im  Blute,  bei  der  Gicht,  ihre  Bindungsweise— 
Gicht  und  Leukamie — die  Absorption  der  Harnsaure  durch  den  Knorpel 
und  ihre  Verhinderung,  Zeit.  exper.  Path.  u.  Ther.  6,  278-299. 

(1911)  Brugsch,  Th.,  and  N.  Masuda:  Ueber  das  Verhalten  des  Diinndarmsaftes 
und  -extraktes,  ferner  des  Extraktes  einiger  Bazillen  (Coli,  Strep tok- 
ken)  gegeniiber  Kasein,  Lecithin,  Amylum.  Ein  Beitrag  zur  funk- 
tionell-diagnostischen  Priifung  der  Faces  auf  Fermente  des  Pankreas, 
Ibid.  8,  617-623;  through  Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  41 
(1911),  295. 

1907a  Brugsch,  Theodor,  and  Alfred  Schittenhelm :  Zur  Stoffwechselpathologie 
der  Gicht.  I.  Mitteilung.  Der  Harnsauregehalt  des  Blutes  bei 
purinfreier  Kost,  Ibid.  4,  438-445. 

1907b  Brugsch,  Theodor,  and  Alfred  Schittenhelm:  Zur  Stoffwechselpathologie 
der  Gicht.  II.  Mitteilung.  Beziehung  zwischen  Blut  und  Harnsaure, 
Ibid.  4,  446-450. 

1907c  Brugsch,  Theodor,  and  Alfred  Schittenhelm:  Zur  Stoffwechselpathologie 
der  Gicht.  III.  Mitteilung.  Der  endogene  und  exogene  Harnsaure- 
und  Purinbasenwerth  bei  der  chronischen  Gicht,  Ibid.  4,  480-531. 

1907d  Brugsch,  Theodor,  and  Alfred  Schittenhelm:  Zur  Stoffwechselpathologie 
der  Gicht.  IV.  Mitteilung.  Ueber  den  Befund  von  Harnsaure  in 
Organen.      Ibid.  4,  532-537. 

1907e  Brugsch,  Theodor,  and  Alfred  Schittenhelm:  Zur  Stoffwechselpathologie 
der  Gicht.  V.  Mitteilung.  Ueber  den  Abbau  von  Glykokoll  und 
Alanin  beim  gesunden  und  gichtkranken  Menschen,  Ibid.  4,  538-550. 

1907f  Brugsch,  Theodor,  and  Alfred  Schittenhelm:  Zur  Stoffwechselpathologie 
der  Gicht.      VI.  Mitteilung.    Pathogenese  der  Gicht,  Ibid.  4,  551,  557. 

1908-9  Brugsch,  Theodor,  and  Alfred  Schittenhelm:  Zur  Stoffwechselpathologie 
der  Gicht.  VII.  Mitteilung.  Das  Verhalten  verfutterter  Purin- 
basen  bei  der  Gicht,  Ibid.  5,  215-226. 

1910  Brugsch,   Theodor,   and  Alfred   Schittenhelm:   Der  Nukleinstoffwechsel 

und  seine  Storungen   (Gicht,  Uratsteindiathese  u.  a.).       Jena,  1910. 

166  pp. 
1905     Buchner,  Eduard,  and  Wilhelm  Antoni:  Existiert  ein  Coenzym  fiir  die 

Zymase  ?  Zeit.  physiol.  Chem.  46,  136-154. 
1908     Buchner,  E.,  and  F.  Klatte:  Ueber  das  Ko-Enzym  des     Hefepresssaftes 

Biochem.  Zeit.  8,  520-557. 


PHOSPHORUS  METABOLISM  603 

1904    Bttchmann,  L.:  Beitrage     zum     Phosphor- Stoffwechsel,  Zeit.  diatet.  u. 

physikal.  Ther.  8,  67-74;  148-160. 
1894     Biilow,  K.:  Ueber  Glycerinphosphorsaure,  Arch.  ges.  Physiol.  57,  89-92. 

1904  Biirger,  Max:  Ueber  Protylin  und  seinen  Wert  als  Nahr-  und  Heilmittel, 

insbesondere  bei  rachitischen  Zustanden  im  Kindesalter,     Therapeut. 

Monatssh.  18,  302-306. 
1913a  Burger,  M.,  and  Beumer:  Zur  Lipoidchemie  des  Blutes,  Berlin,  klin. 

Wochenschr.  50,  112-114. 
1913b  Biirger,  M.,  and  Beumer:  Ueber  die  Phosphatide  der  Erythrocytenstrom- 

ata  bei  Hammel  und  Menschen,  Biochem.  Zeit.  56,  446-456. 

1873  Bunge,  G.:  Ueber  die  Bedeutung  des  Kochsalzes  und  das  Verhalten  der 

Kalisalze  im  menschlichen  Organismus,  Zeit.  Biol.  9,  104-143. 

1874  Bunge,  G. :  Der  Kali-,  Natron-  und  Chlorgehalt  der  Milch,  verglichen  mit 

dem  anderer  Nahrungsmittel  und  des  Gesammtorganismus  der  Sauge- 

thiere,  Ibid.  10,  295-335. 
1876    Bunge,  G.:  Zur  quantitativen  Analyse  des  Blutes,  Ibid.  12,  191-216. 
1885a  Bunge,  G.:  Ueber  die  Assimilation  des  Eisens,  Zeit.  physiol.  Chem.  9, 

49-59. 
1885b  Bunge,  G. :  Analyse  der  anorganischen  Bestandtheile  des  Muskels,  Ibid.  9, 

60-62. 
1886     Bunge,  G.:  Eine  Bemerkung  zur  Theorie  der  Drusenfunction,  Arch.  Anat. 

u.  Physiol.,  physiol.  Abt.,  539,  540. 
1889    Bunge,  G.:  Ueber  die  Aufnahme  des  Eisens    in    den    Organismus    des 

Sauglings,  Zeit.  physiol.  Chem.  13,  399-406. 
1901     Bunge,  G. :  Der  wachsende  Zuckerkonsum  und  seine  Gef  ahren,  Zeit.  Biol. 

41,  155-166. 
1904a  Burian,  Richard:  Zur  Kenntniss  der  Bindung  der  Purinbasen  in  Nuclein- 

sauremolekul,  Ber.  deut.  chem.  Gesell.  37,  I,  708-712. 
1904b  Burian,  Richard:  Zur  Frage  der  Bindung  der  Purinbasen  im  Nuclein- 

sauremolekul,  Zeit.  physiol.  Chem.  42,  297,  298. 

1905  Burian,  Richard:  Ueber  die  oxydative  und  die  vermeintliche  synthet- 

ische  Bildung  von  Harnsaure  in  Rinderleberauszug,  Ibid.  43,  497-531. 
1907a  Burian,  Richard:  Weitere  Beitrage  zur  Kenntnis  der  Diazoaminoverbind- 

ungen  der  Purinbasen,  Ibid.  51,  425-437. 
1907b  Burian,  Richard:  Pyrimidinderivate  aus  Purinbasen,  Ibid.  51,  438-456. 
1897     Burian,  Richard,  and  Heinrich  Schur:     Ueber    die    Nucleinbildung    im 

Saugethierorganismus.      I.    Mitteilung,  Ibid.  23,  55-73. 

1900  Burian,  Richard,   and  Heinrich  Schur:   Ueber  die   Stellung  der  Purin- 

korper  im  menschlichen  Stoffwechsel.       Drei  Untersuchungen,  Arch, 
ges.  Physiol.  80,  241-343. 

1901  Burian,  Richard,  and  Heinrich   Schur:  Ueber  die   Stellung  der  Purin- 

korper  im  menschlichen  Stoffwechsel,  Ibid.  87,  239-354. 
1903     Burian,  Richard,  and  Heinrich  Schur:  Das  quantitative  Verhalten  der 

menschlichen  Harnpurinausscheidung,  Ibid.  94,  273-336. 
1903     Burnet,  James:  On  the  Therapeutic  Value  of  the  Nuclein  Compounds, 

Therapeutic  Gazette   (Detroit)   19,  312,  313. 

1906  Burnett,  E.  A.:  Fattening  Pigs  on  Corn  and  Tankage,  Nebr.  Agr.  Exp. 

Sta.  Bui.  94,  12pp. 
1908    Burnett,  E.  A.:  The  Effect  of  Food  on  Breaking  Strength  of  Bones,  Ibid. 
107,  11-39. 


604  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1910  Burnett,  E.  A.:  The  Effect  of  Food  on  the  Strength,  Size  and  Composi- 
tion of  the  Bones  of  Hogs,  Neb.  Agr.  Exp.  Sta.  Rept.  for  1910,  pp. 
178-208. 

1900  Burow,  Robert:  Der  Lecithingehalt  der  Milch  und  seine  Abhangigkeit 
vom  relativen  Hirngewichte  des  Sauglings,  Zeit.  physiol.  Chem.  30, 
495-507. 

1905  Burow,  Robert:  Beitrage  zur  Entscheidung  der  Frage,  ob  die  Casei'ne 
verschiedener  Tierarten  identisch  sind,  Diss.,  Basel.  28  pp. 

1910  Burow,  Robert:  Ueber  das  Vorkommen  eisenhaltiger  Lipoide  in  der  Milz. 

Vorlaufige  Mitteilung,  Biochem.  Zeit.  25,  165-170. 
1907     Burr,  Anton:  Eigenschaften  und  Zusammensetzung  der  Schweinemilch, 
Milch-Zeitung  36,  565,  566. 

1911  Buslik   and   Goldhaber:    Stoffwechselversuche   mit   Lecithineiweiss   Dr. 

Klopfer  (Glidine),  Zeit.  f.  physikal.  u.     diatet.    Therap.     15,    93-97; 

through  Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  41   (1911),  548. 
1911     Buxton,  B.  H.,  and  A.  H.  Rake:  (Wirkung  der  Verdiinnung  auf  die  Aus- 

flockung  von  Kolloiden),  Jour.  Med.  Research  22,  483-500;   through 

Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  41  (1911),  8,  9. 
1868     Byasson,  H.:  Essai  sur  la  relation  qui  existe  a  l'etat  physiologique  entre 

l'activite  cerebrale  et  la  composition  des  urines,  Paris. 
{1911)  Cagnetto,  G.:  Die  Veranderungen  der  normalen  chemischen  Bestandteile 

des  Knochengewebes  nach  Behandlung  mit   Strontiumsalzen,  Patho- 

logica  3,  137-139;  through  Jahresb.  u.  d.  Fortschr.  d.  Thierchem.  41 

(1911),  361. 
1896     Camerer,   W.    Sr.:   Harnsaure,   Xanthinbasen,   und   Phosphorsaure    im 

menschlichen  Urin,  Zeit.  Biol.  33  (N.  F.  15),  139-155. 
1904     Camerer,  W.  Sr.:  Analysen  vom  menschlichen  Urin,  Ibid.  45  (N.  F.  27), 

1-22. 
1896     Camerer,  W.,  and  Soldner:  Analysen  der  Frauenmilch,  Kuhmilch  und 

Stutenmilch,  Ibid.  33  (N.  F.  15),  535-568. 
1900     Camerer,  W.  Jr.:  Die  chemische  Zusammensetzung  des  Neugeborenen. 

Second  article.      Ibid.  40  (N.  F.  22),  529-534. 
1902a  Camerer,  W.  Jr.:  Die  chemische  Zusammensetzung  des   neugeborenen 

Menschen.      Third  article.      Ibid.  43  (N.  F.  25),  1-12. 
1902b  Camerer,  W.,  Jr.:  Beobachtungen  und  Versuche  liber  die  Ammoniakaus- 

scheidung  im  menschlichen  Urin  mit  Beriicksichtigung  noch  weiterer 

stickstoffhaltiger  Urinbestandteile  und  Bestimmung  der  Aciditat  nach 

Lieblein,  Ibid.  43  (N.  F.  25),  13-45. 
1900     Camerer,   W.   Jr.,   and   Soldner:   Die   chemische   Zusammensetzung   des 

Neugeborenen.       First  article.       Ibid.  39  (N.  F.  21),  173-192. 
1903     Camerer,  W.  Jr.,  and  Soldner:  Die  Aschenbestandteile  des  neugeborenen 

Menschen  und  der  Frauenmilch,  Ibid.  44  (N.  F.  26),  61-77. 
1902     Camerer,  W.,  Soldner  and  Herzog:  Die  chemische  Zusammensetzung  des 

neugeborenen  Menschen,  Ibid.  43  (N.  F.  25),  1-12. 
1909a  Capezzuoli,  Cesare:  Ueber  die    eisenhaltigen    Korper    der    Milz,    Zeit. 

physiol.  Chem.  60,  10-14. 
1909b  Capezzuoli,  Cesare:     Mineralstoffzusammensetzung    der     Knochen    bei 

Osteomalacic,  Biochem.  Zeit.  16,  355,  356. 
1907     Carlier,   Albert:   Experiences  sur  l'influence   du  phosphate   alimentaire 

(Poudre  d'os)  sur  l'elevage  des  gorets,  Annales  de  Gembloux  17,  373- 
377. 


PHOSPHORUS  METABOLISM  605 

1909  Carlier,  Albert :  Experiences  sur  Pemploi  de  la  poudre  d'os  et  de  la  craie 
moulue  dans  Palimentation  des  gorets,  Ibid.  19,  166,  167. 

1892  Carnot,  Adolphe:  Recherche  du  fluor  dans  les  os  modernes  et  les  os 
fossiles,  Compt.  rend.  Acad,  des  sci.  114,  1189-1192. 

1908  Carpiaux,  E. :  Contribution  a  l'etude  de  l'assimilation  du  phosphore  et  de 

la  chaux  pendent  la  vie  embryonaire  du  poussin,  Bui.  Acad,  royal 

Belgique,  CI.  d.  sci.,  1908,  283-295. 
1904     Carre,  P.:  Sur  les  ethers  phosphoriques  de  la  glycerine,  Compt.  rend. 

Acad,  des  sci.  138,  47-49.  . 
1901     Carriere,  G.:  Influence  de  la  lecithine  sur  les  echanges  nutritifs,  Compt. 

rend.  Acad,  des  sci.  133,  314-316. 
1911     Casanova,  Carlo:  (Ueber  Eierlecithin,  iiber  eine  charakteristische  Farb- 

reaktion  und  iiber  Verfalschung  desselben),    Bol.     Chim.     Farm.     50, 

509-513;  through  Zeit.  Unters.  d.  Nahrungs-  u.  Genussmittel  23  (1912), 

695. 
1907    Cathcart,  E.  P.,  and  C.  E.  Fawsitt:  Metabolism  during  Starvation.  Part 

II.      Inorganic.      Jour,  of  Physiol.  36,  27-32. 

1909  Cattaneo,  Cesare:  Ulteriori  ricerche  sul  rapporto  fra  calce  e  magnesio 
nella  ossa  dei  rachitici,  La  pediatria  17,  497,  498;  through  Ch.  Abs.  4 

(1910),  2668. 
1903     Cautru,  Fernand:  Sur  l'innocuite  absolue  de  l'acide  phosphorique,  Bui. 

gener.  de  therapeut.  145,  223-227. 
1904a  Cautru,  Fernand:  Innocuite  et  indications  de  l'acide  phosphorique,  Les 

nouveaux  remedes  20,  535-538. 
1904b  Cautru,  Fernand:  L'acide  phosphorique;  son  innocuite;  ses  principal es 

indications  en  therapeutique,  La  presse  med.,  Paris,  12,  588,  589. 
1904a  Cavazzani,  Emilio :  II  nucleone  nei  centri  nervosi,  Gazzetta  degli  ospedali 

e  delle  cliniche  25,  200,  201. 
1904b  Cavazzani,  Emilio:  Das  Nucleon  im     Ostrea     edulis     (L.),     Centralbl. 

Physiol.  18,  666-668. 
1896    Ceconi,  A.:  Sul  valore  della  determinazione  della  sostanze  organiche 

fosf  orate  nelle  orine  normali  e  pathologische,  7th  Congr.  f .  innere  Med., 

Rom.,  1896;  through  Jahresb.  ii.  d.  Fortschr.  d.  Tierchem.  27  (1897), 

362. 
1898    Ceconi,  A.:  Eliminazione  del  fosforo  organico  durante  l'alta  febbre  e  la 

grave  dispnea,  II  Morgagni,  1898,  No.  3;  through  Centralbl.  innere 

Med.  19,  883. 

1894  Chabrie,  C:  Recherches  sur  les  transformations  chimiques  de  la  sub- 

stance fundamentale  de  cartilage,  Annales  de  chim.  et  de  phys.  333 
(ser.  7,  Vol.  3),  524-563;  also  (briefer  reports)  Compt.  rend.  Acad,  des 
sci.  118  (1894),  1057-1060;  Bui.  Soc.  chim.  66  (1894),  420,  421; 

1895  Chabrie,  C:  Considerations  sur  les  phenomenes  chimiques  de  l'ossifica- 

tion,  Compt.  rend.  Acad,  des  sci.  120,  1226-1228. 

1911  Chamberlain,  Weston  P.:  The  Eradication  of  Beriberi  from  the  Philippine 
(Native)  Scouts  by  Means  of  a  Simple  Change  in  their  Dietary,  Phil- 
ippine Jour,  of  Sci.,  B.,  6,  133-146. 

1910  Chamberlain,  W.  P.,  H.  D.  Bloombergh  and  E.  D.  Kilbourne:  A  Study  of 
the  Influence  of  Rice  Diet  and  of  Inanition  on  the  production  of  Multi- 
ple Neuritis  of  Fowls  and  the  Bearing  thereof  on  the  Etiology  of 
Beriberi,  Ibid.  6,  177-208. 


606  OHIO  EXPEEIMENT  STATION:  TECHNICAL  BUL.  5 

1910  Chamberlain,  W.  P.,  and  E.  B.  Vedder:  A  Contribution  to  the  Etiology  of 

Beriberi.  Ibid  6,  251-258. 

1911  Chamberlain,  W.  P.,  and  E.  B.  Vedder:  A  Second  Contribution  to  the 

Etiology  of  Beriberi,  Ibid.  6,  395-404. 

1912  Chamberlain,  Weston  P.,  Edward  B.  Vedder,  and  Robert  R.  Williams:  A 

Third  Contribution  to  the  Etiology  of  Beriberi,  Ibid.  7,  39-52. 
1905     Charrin,   A.:  Action   des   matieres  minerales   sur   les   echanges   et   la 
resistance  de  l'organisme,  Compt.  rend.  Soc.  de  biol.  59,  112-114. 

1896  Charrin,  A.,  and  Desgrez:  Action  des  solutions  mineralisees  sur  l'organ- 

isme, Ibid.  48,  805,  806. 
1895     Chatin,  A.,  and  A.  Muntz:  Existence  du  phosphore  en  proportion  notable 

dans  les  huitres,  Comp.  rend.  Acad,  des  sci.  120,  1095-1097. 
1908    Chavan,  Paul:  Influence  de  la  fumure  phosphatee  et  potassique  sur  la 

composition  chimique  du  fourrage  de     prairie     naturelle,     Annuaire 

agric.  de  la  Suisse  9  (22  of  Ger.  edition),  193-206. 
1886    Chevalier,  Josephine:  Chemische    Untersuchung    der    Nervensubstanz, 

Zeit.  physiol.  Chem.  10,  97-105. 

1897  Chittenden,  R.  H.:  The  Protagon  of  the  Brain,  Proc.  Amer.  Physiol* 

Soc,  Science  5  (N.  S.),  909. 
1885     Chittenden,  R.  H.,  and  H.  M.  Painter:  Casein  and  its  Primary  Cleavage 
Products,   also    Caseoses,   Casein    Dyspeptone    and    Casein    Peptone, 
Studies  from  Lab.  of  Physiol.  Chem.,  Yale  Univ.,  II,  156-199;   III, 
66-105. 

1842  Chossat,  Charles:  Note  sur  le  systeme  osseux,  Compt.  rend.  Acad,  des 

sci.  14,  451-454. 

1843  Chossat,  Charles:  Recherches  experimentales  sur  l'inanition,  Mem.  pre- 

sented a  l'Acad.  des  sci.  de  lTnst.  de  France  8  (ser.  2),  438-640. 
1910     Choumowa-Sieber,  N.  O.:  Influence  exercee  par  l'alcool  sur  la  teneur  des 

organes  des  animaux  en  phosphatides,  Archives  des  sci.  biol.,  l'Inst. 

imper.  de  med.  exper.  a,  St.  Petersbourg  15,  373-392.      56  refs. 
1910     Ciaccio,  C:  Contributo  alia  distribuzione  ed  alia  fisio-patologia  cellulare 

dei  lipoidi,  Arch.  f.  Zellf.  5,  235-363;  through  Zentralbl.  Biochem.  u. 

Biophys.  11  (N.  F.  2)  (1910-11),  322,  323. 
1910     Ciuffini,  Publio:  II  ricambio  del  calcio,  del  magnesio,  e  dell'  acido  fosfor- 

ico  nell'  uomo  sano  e  nel  gottoso,  II  Policlinico,  sezione  med.  17,  156- 

180. 
1901a    Claude,  H.,  and  A.  Zaky:  Le  lecithine  dans  la  tuberculose  (Note  prelim- 

inaire),   Compt.   rend.    Soc.   de   biol.  53,   821-823;    also   Compt.   rend. 

Acad,  des  sci.  133,  486-488. 
1901b  Claude,  H.,  and  A.  Zaky:  La  lecithine  dans  la  tuberculose,  La  Presse 

med.  9,  II,  173-178. 
1901c  Claude,  H.,  and  A.  Zaky:  La  lecithine  dans  la  tuberculose.  Gazette  des 

hospitaux  civils  et  militaires  74,  1084. 
1910     Clementi,   A.:    Intorno    all'azione   della   lipasi   pancreatica    ed   enterica 

sulla   lecitina,  Archivio   di  fisiologia   8,   399-408;    through  Zentralbl. 

Biochem.  u.  Biophys.  11  (1911),  435. 
1901     Cocchi,  A.:   Sopra  il  nucleoproteide  della  placenta  umana,  Lo   Speri- 

mentale  55,  503-508. 
1908     Cohen,  L.  J.,  and  William  J.  Gies:  A  Study  of  "Protagon"  Prepared  by 

the  Wilson  and  Cramer  Method,  Proc.  Soc.  Exp.  Biol,  and  Med.  5, 

97-100. 


PHOSPHORUS  METABOLISM  607 

1889  Cohn,  Conrad:  Ueber  den  Einfluss  der  Caries  auf  die  chemische  Zusam- 
mensetzung  des  Zahnbeines,  Inaug.  Diss.,  Berlin.    25  pp. 

1907  Cohn,  Michal:  Kalk,  Phosphor  und  Stickstoff  im  Kindergehirn,  Deut. 

med.  Wochenschr.  33,  1987-1991. 
1913     Cohn,   R.:   Ueber   den  "Riickgang"   der  Lecithinphosphorsaure,   Chem. 

Zeitung  37,  581-583. 
1870     Cohn,  Wilhelm:  Ueber  die  Verwendung    des    basisch    phosphorsauren 

Kalkes  bei  der  Aufzucht  von  Vieh,  Annalen  der  landwirtsch.  Woch- 

enbl.,  Konig.  Preussisch.  Statten,  pp.  431-433. 
1882     Cohnheim,  Julius:  Vorlesungen  iiber  allgemeine  Pathologie;  ein  Handbuch 

fiir  Aerzte  und  Studierende.  2nd  edition.       Berlin.  2  Vols. 

1908  Cohnheim,  Otto:  Die  Physiologie  der  Verdauung  und  Ernahrung.       23 

Vorlesungen  fiir  Studierende  und  Aerzte.       Berlin  and  Wien.  479  pp. 

1902  Colombet,  Andre:  De  la  medication  arsenio-phosphoree  organique  dans  le 

traitement  de  la  tuberculose  pulmonaire  chronique,  These  de  Paris. 

1903  Concetti,  Luigi:  La  cura  del  fosforo  nel  rachitismo,  Rivista  di  clinica 

pediat.      1,  24-37. 

1909  Cook,  F.  C:  Metabolism  of  Organic  and  Inorganic  Phosphorus,  U.  S. 

Dept.  Agr.,  Bur.  Chem.,  Bui.  No.  123. 

1911  Cooper,  E.  A.,  and  Casimir  Funk :  Experiments  on  the  Causation  of  Beri- 

beri, Lancet  1911,  II,  1266,  1267. 

1903  Coriat,  Isador  H.:  A  Contribution  to  the  Chemistry  of  Nerve  Degenera- 
tion in  General  Paralysis  and  Other  Mental  Disorders,  Amer  Jour. 
Insan.  59,  393-416. 

1904a  Coriat,  Isador  H.:  The  Production  of  Cholin  from  Lecithin  and  Brain- 
Tissue,  Amer.  Jour.  Physiol.  12,  353-362. 

1904b  Coriat,  Isador  H.:  Chemical  Findings  in  Cerebrospinal  Fluid,  Amer.  Jour. 
Insan.  60,  733-761. 

1905  Coriat,  Isador  H. :  A  Review  of  Some  Recent  Literature  on  the  Chemistry 

of  the  Central  Nervous  System,  Jour,  of  Comp.  Neurol.  15,  148-159. 

1912  Corper,  Harry  J.:  Chemistry  of  the  Dog's  Spleen,  Jour.  Biol.  Chem.  11, 

27-35. 
1912     Costantino,  A.:  Beitrage  zur  Muskelchemie.      II.  Ueber  den  Gehalt  der 

glatten  und  quergestreiften   Saugetiermuskeln   an   organischem  und 

anorganischem  Phosphor,  Biochem.  Zeit.  43,  165-180. 
1834     Couerbe,  J.  P.:  Du  cerveau,  considere  sous  le  point  de  vue  chimique  et 

physiologique,  Annales  de  chim.  et  de  phys.  152  (ser.  2,  56),  160-193. 
1901     Coulombe,  Edmond:  La  lecithine  de  l'oeuf,  son  emploi  therapeutique,  These 

de  Paris.  56  refs.    70  pp. 
1891     Courant,  Georg:  Ueber  die  Reaktion  der  Kuh-    und    Frauenmilch    und 

ihre  Beziehungen  zur  Reaktion  des  Caseins  und  der  Phosphate,  Arch. 

ges.  Physiol.  50,  109-165. 
1903     Courtial,  C:  Etude  de  la  lecithine  et  des  jaunes  d'oeufs  industriels, 

These  Montpellier;  through  Jahresb.  u.  d.  Fortschr.  d.  Thierchem.  33 

(1903),  870. 
1903     Cousin,  H.:  Sur  les  acides  gras  de  la  lecithine  de  l'oeuf,  Compt.  rend. 

Soc.  de  biol.  55,  913-915;  also  Compt.  rend.  Acad,  des  sci.  137,  68-70. 

1906  Cousin,  H.:  Sur  les  acides  gras  de  la  cephaline,  Compt.  rend.  Soc.  de 

biol.  61,  23-25;  also  Jour,  de  pharm.  et  de  chim.  24  (1906),  101-108. 


608  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1904  Cramer,  W. :  On  Protagon,  Cholin,  and  Neurin,  Jour,  of  Physiol.  31,  30- 

37. 
1910     Crawford,  Albert  C:  A  Poisonous  Principle  in  Certain  Cottonseed  Meals, 

Jour,  of  Pharm.  and  Exp.  Ther.  1,  519-548. 
1891     Cremer,  Max:  Demonstration  eines     mit    kalkarmer     Nahrung    unter 

Zugabe  von  Strontiumphosphate  ernahrten  jungen  Hundes,  Sitzungs- 

ber  d.  Gesell.  f.  Morph.  u.  Phys.  in  Mtinchen  1891,  124-126. 

1910  Cronheim,  J.  and  W.:  Weitere  Untersuchungen  liber  die  Bedeutung  des 

Lecithins  fur  den  Stoffwechsel  des  Sauglings,  Zeit.  physikal.  u.  diatet. 

Ther.  14,  257-267. 
1911a  Cronheim,  J.  and  W.:  Erwiderung,  Ibid.  15,  101. 
1911b  Cronheim,  J.  and  W.:  Schlusswort,  Ibid.  15,  359. 
1912     Cronheim,  W. :  Die  Bedeutung  des  Lecithins  im  Stoffwechsel  des  Erwach- 

senen,  Ibid.  16,  262-272. 

1900  Cronheim,  W.,  and  Erich  MUller:  Zur  Kenntnis  der  Bedeutung  des  organ- 

isch  gebundenen  Phosphors  fur  den  Stoffwechsel  des  Kindes,  Jahrb. 
Kinderheilk.  52,  360-364. 

1902  Cronheim,  W.,  and  Erich  MUller:  Versuche  iiber  den  Stoff-  und  Kraft- 

wechsel  des  Sauglings  mit  besonderer  Beriicksichtigung  des  organisch 
gebundenen  Phosphors,  Zeit.  diatet.  u.  physikal.  Ther.6,  25-44;  92-114. 

1903  Cronheim,  W.,  and  Erich  MUller:  Untersuchungen  uber  den  Einnuss  der 

Sterilisation   der   Milch   auf   den   Stoffwechsel    des   Sauglings   unter 

besonderer  Berucksichtigung  der  Knochenbildung,  Jahrb.  Kinderheilk. 

57,  45-63. 
1908     Cronheim,  W.,  and  Erich  MUller:  Stoffwechselversuche  an  gesunden  und 

rachitischen  Kindern  mit  besonderer  BerUcksichtigung  des  Mineral- 

stoffwechsels,  Biochem.  Zeit.  9,  76-126. 
1895     Curatulo,  G.  E.,  and  L.  Tarulli:  Einnuss  der  Abtragung  der  Eierstoke 

auf  den  Stoffwechsel,  Centralbl.  Physiol.  9,  149-152;  through  Jahresb. 

u.  d.  Fortschr.  d.  Thierchem.  25  (1895),  498,  499. 

1911  Curschmann,  H.:  Ueber  Osteomalacia  senilis  und  tarda,  Med.  Kiinik  7, 

1565-1571. 

1912  Czernoruzky,  M.  W.:  (Ueber  die  Wirkung  der  Nukleinsaure  auf  die  fer- 

mentativen  Prozesse  im  Tierorganismus),  Wratschebnaja  Gazeta, 
1912,  No.  14  and  15;  through  Zeit.  Immunitatsforschung,  II.  Teil, 
Referate  5  (1912),  328.       (See  also  Tschernoruzki,  1911.) 

1866  Dahnhardt,  C:  Zur  Chemie  der  Lymphe,  Arch.  path.  Anat.  u.  Physiol.  37, 
55-67. 

1870  Dahnhardt,  C:  Zur  Caseinbildung  in  der  MilchdrUse,  Arch.  ges.  Physiol. 
3,  586-598. 

1913a  Dakin,  H.  D.,  and  H.  W.  Dudley:  The  Racemization  of  Proteins  and  their 
Derivatives  Resulting  from  Tautomeric  Change.  II.  The  Racemiza- 
tion of  Casein,  Jour.  Biol.  Chem.  15,  263-269. 

1913b  Dakin,  H.  D.,  and  H.  W.  Dudley:  The  Action  of  Enzymes  on  Racemized 
Proteins  and  their  Fate  in  the  Animal  Body,  Ibid.  15,  271-276. 

1901  Dalmastri :  II  zicambio  dell'  azoto  e  del  f osf oro  durante  la  cura  antirabica 

Bollett.  delle  sci.  med.  di  Bologna,  Apr.  1901,  No.  4;  through  Jahresb. 
u.  d.  Fortschr.  d.  Thierchem.    31  (1901),  766,  767. 

1905  Dambre,  L.  A.:  Contribution  a  Petude  de  la  medication  phosphoree.  La 

phytine,  Thesis,  Toulouse.  64  pp.  Through  Biochem.  Centralbl.  5 
(1906-7),  133. 


PHOSPHORUS  METABOLISM  609 

1911a  Daniel-Brunet,  A.,  and  C.  Rolland:  De  l'influence  du  sexe  et  de  la  castra- 
tion sur  la  quantite  des  lipoi'des  de  la  bile  chez  les  bovides,  Compt. 
rend.  Acad,  des  sci.  153,  214,  215. 

1911b  Daniel-Brunet,  A.,  and  C.  Rolland:  Contribution  a  l'etude  chimique  et 
physiologique  de  la  glande  hepatique  des  bovides,  Ibid.  153,  900-902. 

1895a  Danilewsky,  B.:  Ueber  die  Blutbildende  Eigenschaft  der  Milz  und  des 
Knochenmarks,  Arch.  ges.  Physiol.  61,  264-274. 

1895b  Danilewsky,  B.:  De  l'innuence  de  la  lecithine  sur  la  croissance  et  multi- 
plication des  organismes,  Compt.  rend.  Acad,  des  sci.  121,  1167-1170; 
also  Westnik  mediciny  (1896)  1,  1  and  269-285  (Russian). 

1896  Danilewsky,  B.:  De  l'influence  de  la  lecithine    sur    la    croissance    des 

animaux  a  sang  chaud,  Compt.  rend.  Acad,  des  sci.  123,  195-198. 

1897  Danilewsky,  B.:  Influence  des  lecithines  sur  le  croissance,  Compt.  rend. 

Soc.  de  biol.  49,  475,  476. 
1902     Dapper,  Max:  Ueber  Fleischmast  beim  Menschen,  Inaug.  Diss.,  Marburg. 
1871     Decaisne,  E. :  Des  modifications  que  subit  le  lait  de  f  emme  par  suite  d'une 

alimentation  insuffisante.       Observations  recueillies  pendant  le  siege 

de  Paris,  Gaz.  med.  de  Paris,  p.  317. 

1898  Decroly,  O.: Etude  de  Taction  des  toxines  et  antitoxines  sur  la  nutrition 

generale,  Archives  internat.  de  pharmacodynamic  4,  385-489. 
1911     Delaini,  G.:  Sul  comportamento  degli  ipofosfiti  nell-  organismo  animale, 
Archivio  de  fisiologia  9,  329-340. 

1899  Delcourt,  Albert :  Le  rachitisme,  Ann.  Soc.  des  sci.  med.  et  naturelles  de 

Bruxelles  8,  1-104. 
1909    Delille,  Arthur:  L'hypophyse  et  la  medication    hypophysaire.     (Etude 
experimentale  et  clinique.)      These  de  Paris.      366  pp.,  14  pages  refs. 

1905  Dennstedt,  M.,  and  Th.  Rumpf :  Weitere  Untersuchungen  uber  die  chem- 

ische  Zusammensetzung  des  Blutes  und  verschiedener  menschlicher 
Organe  in  der  Norm  und  in  Krankheiten,  Zeit.  klin.  Med.  58,  84-162. 

1906  Desgrez,  A.,  and  Bl.  Guende:  Influence  de  l'acide  phosphorique,  des  phos- 

phates mono  et  disodiques  sur  les  echanges  nutritifs,  Compt.  rend. 
Acad,  des  sci.  142,  1440-1442. 

1907  Desgrez,  A.,  and  J.  Posen:  Sur  la  determination  de  la  molecule  elaboree 

moyenne  et  ses  variations,  dans  l'organism  animal,  sous  1'influence  des 
composes  mineraux  du  phosphore,  Compt.  rend.  Soc.  de  biol.  63,  455- 
457. 

1900  Desgrez,  A.,  and  A.  Zaky:  De  l'influence  des  lecithines  sur  les  echanges 

nutritifs,  Ibid.  52,  794,  795. 
1901a  Desgrez,  A.,  and  A.  Zaky:  Influence  des  lecithines  de  l'oeuf  sur  les 

echanges  nutritifs,  Ibid.  53,  647-649;  also  Compt.  rend.  Acad,  des  sci. 

132, 1512-1514. 
1901b  Desgrez,  A.,  and  A.  Zaky:  Influence  de  le  lecithine  sur  l'elimination  de 

l'acide  urique,  Compt.  rend.  Soc.  de  biol.  53,  830-832. 
1902a  Desgrez,  A.,  and  A.  Zaky:  De  l'influence  des  lecithines  sur  le  developp- 

ment  du  squelette  et  du  tissu  nerveux,  Ibid.  54,  501-504;  also  Compt. 

rend.  Acad,  des  sci.  134,  1166-1168. 
1902b  Desgrez,  A.,  and  A.  Zaky:  Analyse  du  mode  d'action  des  lecithines  sur 

l'organisme  animal,  Compt.  rend.  Soc.  de  biol.  54,  730,  731;  also  Compt. 

rend.  Acad,  des  sci.  134,  1522,  1523. 


610  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1902c  Desgrez,  A.,  and  A.  Zaky:  Etude  de  l'influence  des  lecithines  sur  l'organ- 
isme  animal,  Jour,  de  physiol.  et  de  path,  gener.  4,  662-671. 

1904a  Desgrez,  A.,  and  A.  Zaky:  De  Tinfluence  comparee  des  composes  organ- 
iques  phosphores  sur  la  nutrition,  sur  le  developpment  et  la  composi- 
tion des  tissus  animaux,  Revue  Soc.  sci.  hygiene  alimentaire  (1904) 
825-831. 

1904b  Desgrez,  A.,  and  A.  Zaky:  De  l'influence  comparee  des  composes  des 
organiques  phosphores  sur  la  nutrition,  Compt.  rend.  Soc.  de  biol.  57, 
392-395;  440-443. 

1904c  Desgrez,  A.,  and  A.  Zaky:  Influence  comparee  de  quelques  composes 
organiques  du  phosphore  sur  la  nutrition  et  la  developpement  des 
animaux,  Compt.  rend.  Acad,  des  sci.  139,  819-821. 

1905  Desgrez,  A.,  and  A.  Zaky:  Etude  de  l'influence  de  quelques  composes 
organiques  phosphores  sur  l'organisme  animal,  Jour,  de  physiol.  et  de 
path,  gener.  7,  213-220. 

1898  Deucher,  P.:  Stoffwechseluntersuchungen  bei  Verschluss  des  Ductus 
pancreaticus,  Correspondenzblatt  fur  Schweizer  Aerzte  28,  321-329; 
361-366. 

1909  Dhere,  Ch.,  and  H.  Maurice:  Influence  de  l'age  sur  la  quantite  et  la 

repartition  chimique  du  phosphore  contenu  dans  les  nerfs,  Compt.  rend. 
Acad,  des  sci.  148,  1124,  1125. 

1910  Dhere,  Ch.,  and  H.  Maurice:  Influence  de  l'age  sur  la  quantite  et  la 

repartition  chimique  du  phosphore  contenu  dans  la  rate,  Compt.  rend. 

Soc.  de  biol.  69,  311,  312. 
1867a  Diaconow,  C:  Ueber  die    phosphorhaltigen    Korper    der    Hiihner    und 

Storeier,  Hoppe-Seyler's  med.  -chem.  Untersuchungen  (1867),  221-227. 
1867b  Diaconow,  C:  Ueber  das  Lecithin,  Ibid.,  405-411. 
1868a  Diaconow,  C:  Das  Lecithin  im  Gehirn,  Centralbl.  f.  med.  Wissensch.    6, 

97-99. 
1868b  Diaconow,  C:  Ueber  die  chemische  Constitution  des  Lecithins,  Ibid.  6, 

2,  3;  434,  435. 

1908  Dibbelt,   W.:  Die  Pathogenese  der  Rhachitis.  I.  Arbeiten  a.  d.  Gebiete  d. 

path.  Anat.  u.  Bakteriol.  aus  dem  path.  anat.  Inst.  Tubingen  6,  670- 
709.      50  refs. 

1909  Dibbelt,  W.:  Die  Pathogenese  der  Rhachitis.  II.  Ibid.  7,  144-214.  31  refs. 
1910a  Dibbelt,  W.:  Weitere  Beitrage  zur  Pathogenese  der  Rhachitis,  Verhandl. 

deut.  path.  Gesell.  14,  294-299. 
1910b  Dibbelt,  W. :  Die  Bedeutung  der  Kalksalze  f iir  die  Schwangerschaf ts-  und 
Stillperiode   und   der   Einflusz   einer   negativen   Kalkbilanz   auf   den 
miitterlichen  und  kindlichen  Organismus,  Ziegler's  Beitrage  48,  147- 
169. 

1911  Dibbelt,  W.:  Die  experimentelle  Osteomalacic  und  ihre  Heilung.  Zugleich 

ein  kritischer  Beitrag     zur     Histogenese     der     Skeletterkrankungen, 
Arbeiten  aus  dem  path.  Inst.  Tubingen  7,  559-588. 

1912  Dibbelt,  W.:  Neue  experimentelle  Untersuchungen  iiber  die  Pathogenese 

der  Rachitis,  Deut.  med.  Wochenschr.  38,  I,  316-318. 

1913  Dibbelt,  W.:  Die  Aetiologie  der  Rachitis  und  der  Kalkstoffwechsel,  Ibid. 

39,  551,  552. 
1909     Diesing,  Ernst:  Beitrag  zur  Kenntniss  der  Funktion  der  Stoffwechsel- 
driisen,  Zentralbl.  ges.  Physiol,  u.  Path.  d.  Stoffwechs.  n.  ser.  4,  209- 
216. 


PHOSPHORUS  METABOLISM  611 

1913    Diesing:  Beitrag  zur  Aetiologie  der  Rachitis,  Deut.  med.  Wochenschr. 

39,  552. 
1909     Dietrich,  M.:  Ueber  phosphorhaltige  Caseinpeptone,  Biochem.  Zeit.  22, 

120-130. 
1902     Dietrich,  Th.:  Getrocknete  Biertreber,  Landwirtsch.  Versuch.  Stat.  56, 

207-256;  257-262. 
1904    Diffloth,  Paul:  Du  role  de  quelques  agents  physiques  et  chimiques  dans 

l'insolubilisation  des  phosphates  du  lait,  Bui.  des  sci.  Pharmacol.  10, 

273-279. 
1900    Dmitriewski,  K.:  Stoffwechsel  bei  wiederholter  Injection  von  bacteriellen 

Giften,  Russ.  Arch.  f.  Path.,  klin.  Med.  u.  Bacteriol.  9,  375-391;  through 

Jahresb.  u.  d.  Fortschr.  d.  Thierchem.  30  (1900),  778,  779. 
1911    Dobrowolsku,  W.  M.:   (Die  Behandlung  der     Hunde     mit     erweichten 

Knochen),  Verhandl.  d.  Gesellsch.  russ.  Aerzte  1911,  195;   through 

Jahresb.  u.  d.  Fortschr.  d.  Thierchem.  41  (1911),  360. 
1906    Doctor,  Earl:  Ueber  die  chemische  Reaction  des  Harnes  mit  Riicksicht 

auf  die  Phosphaturie,  "Urologia"  a  Budapesti  Orvosi  Ujsag  melleklete, 

Budapest,  4,  88,  89;  through  Pester  med.-  chir.  Presse  43  (1907). 
1885    Dogiel,  A.:  Einiges  uber  die    Eiweisskorper    der    Frauen-      und      der 

Kuhmilch,  Zeit.  physiol.  Chem.  9,  591-615. 
1913     Dohrn,  Max:  Beitrag  zum  Nucleinstoffwechsel,  Ibid.  86,  130-136. 
1876    Domenico,  Pecile:  Guanin  im  Schweineharn,    Annalen    der    Chem.    u. 

Pharm.  183,  141-144. 
(1911)  Donath,  Hedwig:  Ueber  die  therapeutische  Anwendung  des  Phytinum 

liquidum  in  der  Kinderheilkunde,  Wien.  klin.  Wochenschr.  24,  1192-1197; 

through  Jahresb.  u.  d.  Fortschr.  d.  Thierchem.  41  (1911),  842. 
1903a  Donath,  Julius:    Die  Behandlung    der    progressiven    Paralyse,    sowie 

toxischer  und  infectioser  Psychosen  mit  Salzinfusionen,  Allgem.  Zeit. 

Psychiat.  u.  psychischgerichtl.  Med.  60,  583-605. 
1903b  Donath,  Julius:  Das  Vorkommen  und  die  Bedeutung  des  Cholins  in  der 

Cerebrospinalfliissigkeit  bei  Epilepsie  und  organischen  Erkrankungen 

des  nervensystems,  nebst  weiteren  Beitragen  zur  Chemie  derselben, 

Zeit.  physiol.  Chem.  39,  526-544. 
1904    Donath,  Julius:  Die  Phosphorsauregehalt  der   Cerebrospinalfliissigkeit 
bei  verschiedenen  Nervenkrankheiten,  Ibid.  42,  141-148. 

1909  Donath,  Julius:  Die  Behandlung  der  progressiven  allgemeinen  Paralyse 

mittels  Nuklein-Injektionen,  Wien.  klin.  Wochenschr.  22,  1289-1292. 
1890    Doremus,  Ch.  A:  Ueber  Elefantenmilch,  Molkereizeitung  p.  67;  through 

Jahresb.  u.  d.  Forschr.  d.  Thierchem.  20  (1890),  147. 
1904    Dorn,  Franz:  Beitrag  zur  Phosphorbehandlung  mit  besonderer  Beriick- 

sichtigung    der    Wirkung    des    Protylin-Roche    bei    Rachitis    und 

Skrofulose,  Deut.  Aerzte-Zeitung,  265-270. 

1910  Dornic  and  Daire:  Contribution  a  l'etude  de  la  lecithine  du  lait,  Annales 

des  falsifications  3,  533-538. 
1912    Doyon,  M.:  Extraction  de  l'antithrombine  des  testicules  et  de  l'intestin, 

Compt.  rend.  Soc.  de  biol.  72,  925,  926. 
1912    Doyon,  M.,  and  P.  Dubrulle:  Formation  d'une  substance  anticoagulante 

phosphoree  sous  l'influence  de  l'autodigestion  de  l'intestin,  Ibid.  73, 

546. 


612  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1912  Doyon,  M.,  P.  Dubrulle  and  F.  Sarvonat:  Digestion  pepsique  de  la  nucleo- 

proteide  extraite  de  l'intestin.  Comparaison  du  pouvoie  anticoagu- 
lant de  la  substance  initiale  et  du  residu,  Ibid.  73,  720. 

1912a  Doyon,  M.,  and  F.  Sarvonat:  Proprietes  anticoagulantes  de  l'acide 
nucleinique  extrait  de  l'intestin,  Ibid.  73,  546,  547. 

1912b  Doyon,  M.,  and  F.  Sarvonat:  Proprietes  anticoagulantes  des  acides 
nucleiniques  d'origine  animale  et  vegetale,  Ibid.  73,  619. 

1912c  Doyon,  M.,  and  F.  Sarvonat:  Proprietes  anticoagulantes  des  acides 
thymo-nucleinique  et  thymique,  Ibid.  73,  644. 

1913a  Doyon,  M.,  and  F.  Sarvonat:  Proprietes  anticoagulantes  de  l'acide 
nucleinique  extrait  des  globules  du  sang  des  oiseaux,  Ibid.  74,  312. 

1913b  Doyon,  M.,  and  F.  Sarvonat:  Action  anticoagulante  de  l'hematogene, 
Ibid.  74,  368,  369. 

1913c  Doyon,  M.,  and  F.  Sarvonat:  Action  comparee  des  divers  phosphates  sur 
la  coagulation  du  sang,  Ibid.  74,  460. 

1913d  Doyon,  M.,  and  F.  Sarvonat :  Action  comparee  du  nucleinate  de  soude  sur 
la  coagulation  du  sang  et  sur  la  coagulation  du  lait,  Ibid.  74,  765,  766. 

1913e  Doyon,  M.,  and  F.  Sarvonat:  Pouvoir  glycolytique  du  sang  preleve  pen- 
dent l'intoxication  provoquee  par  les  peptones,  Ibid.,  74,  779. 

1913f  Doyon,  M.,  and  F.  Sarvonat:  Nucleinate  de  soude  et  pouvoir  coagulant 
du  serum,  Ibid.  74,  872-874. 

1911  Draeger,  Ewald:  Ueber  die  Assimilation  und  Verwertung  verschieden- 
er  Kalkphosphatpraparate  im  Tierkorper,  Diss.  Rostock  1911,  51  pp. 
Through  Jahresb.  u.  d.  Fortschr.  d.  Thierchem.  41  (1911),  893. 

1886  Drechsel,  E.:  Ueber  einen  neuen,  schwefel-  und  phosphorhaltigen 
Bestandtheil  der  Leber,  Jour,  prakt.  Chem.  33,  425-432. 

1913  Drbge,  Karl:  Ueber  Veranderungen  in  der  chemischen  Konstitution  des 

Tierkorpers  nach  Exstirpation  der  Milz,  der  Hoden  und  des  Schild- 
drusenapparates,  Arch.  ges.  Physiol.  152,  437-477. 

1914  Du  Bois,  Gaston:  The  Chemistry  and  Properties  of  Glycerophosphates 

(Glycerinophosphates)  Jour,  of  Ind.  and  Engin.  Chem.  6, 122-128. 
1884a  Duclaux,  Emile:  Sur  les  matieres  albuminoides  du  lait,  Compt.  rend. 

Acad,  des  sci.  98,  373-375. 
1884b  Duclaux,  Emile:  Sur  la  constitution  du  lait,  Ibid,  98,  438-441. 
1884c  Duclaux,  Emile:  Action  de  la  presure  sur  le  lait,  Ibid.  98,  526,  528. 

1893  Duclaux,  Emile:  Sur  les  phosphates  du  lait,  Annales  de  llnst.  Pasteur 

7,  2-17. 

1894  Duclaux,  Emile:  Le  lait,  Paris.  371  pp. 

1905    von  During,  E.:  Ueber  Phosphaturie,  Med.  Klinik,  Berlin,  1,  513-516. 
1903-4  Dunham,  Edward  K.:  The  Lecithin  Content  of  Fatty  Extracts  from  the 

Kidney,  Proc.  Soc.  Exp.  Biol,  and  Med.,  New  York,  1,  39-41. 
1904-5  Dunham,  Edward  K. :  Further  Observations  upon  the  Phosphorized  Fats 

in  Extracts  of  the  Kidney,  Ibid.  2,  63,  64. 
1905-6  Dunham,  Edward    K.:      Analogies    between    the    Phosphorized    Fats 

Obtained  from  the  Brain  and  Kidney,  Ibid.  3,  67-70. 
1908    Dunham,  Edward  K. :  The  Isolation  of  Carnaubic  Acid  from  Beef  Kidney, 

Jour.  Biol.  Chem.  4,  297-299. 
1910    Dunham,  Edward  K.,  and  C.  A.  Jacobson:  Ueber  Carnaubon:  Ein  glycer- 

infreies  Phosphatid,  lecithinahnlich  konstituiert  mit  Galaktos  als  Kern, 

Zeit.  physiol.  Chem.  64,  302-315. 


PHOSPHORUS  METABOLISM  613 

1897-8  Dunlap,  J.  C,  D.  Noel  Paton  and  R.  Stockman :  On  the  Influence  of  Mus- 
cular Exercise,  Sweating  and  Massage,  on  the  Metabolism,  Jour,  of 
Physiol.  22,  68-91. 

1913  Durlach,  Ernst.:  Untersuchungen  liber  die  Bedeutung  des  Phosphors  in 
der  Nahrung  wachsender  Hunde,  Arch.  exp.  Path.  u.  Pharm.  71,  210- 
250. 

1869  Dusart,  L.:  Recherches  experimentales  sur  les  proprietes  physiologiques 

et  therapeutiques  du  phosphate  de  chaux,  Archives  gener.  de  med.,  6th 
ser.,  14,  670-695. 

1870  Dusart,  L.:  Recherches  experimentales  sur  les  proprietes  physiologiques 

et  therapeutiques  du  phosphate  de  chaux,  Ibid.  15,  66-84. 
1913     Eckles,  C.  H.,  and  R.  H.  Shaw:  The  Influence  of  the  Stage  of  Lactation 
on  the  Composition  and  Properties  of  Milk,  U.  S.  Dept.  Agr.,  Bur. 
Animal  Ind.,  Bui.  155,  88  pp. 

1912  Edie,  E.  S.,  W.  H.  Evans,  B.  Moore,  G.  C.  E.  Simpson,  and  A.  Webster: 

The  Anti-Neuritic  Bases  of  Vegetable  Origin  in  Relationship  to  Beri- 
beri, with  a  method  of  Isolation  of  Torulin,  the  Anti-Neuritic  Base 
of  Yeast,  Biochemical  Jour.  6,  234-242. 

1911  Edie,  E.  S.,  and  G.  C.  E.  Simpson:  The  Preparation  of  Various  Foodstuffs 
(Especially  Wheat  and  Rice) :  Its  Effects  on  their  Content  of  Organic 
Phosphorus  Compounds  and  its  Relation  to  Disease,  British  Med. 
Jour.  1,  1421,  1422. 

1891  Edkins,  J.  Sydney:  The  Changes  Produced  in  Casein  by  the  Action  of 
Pancreatic  and  Rennet  Extracts,  Jour,  of  Physiol.  12,  193-219. 

1878  Edlefsen,  G.:  Ueber  das  Verhaltniss  der  Phosphorsaure  zum  Stickstoff 
im  Urin,  Centralbl.  med.  Wissensch.  16,  513-523. 

1880  Edlefsen,  G.:   Ueber  die  Ableitung  der  specifischen   Gallenbestandteile 

und  des  Glycogens  neben  Harnstoff  aus  der  Formel  des  Hamoglobins 
und  iiber  den  relativen  Wert  der  aus  den  Blutkorperchen  abzuleitend- 
en  Phosphorsaure  im  Urin,  Ibid.  18,  657-661;  675-677;  691-696. 

1881  Edlefsen,   G.:   Verhaltnis   der   Phosphorsaure   zum   Stickstoff  im   Urin, 

Deut.  Arch.  klin.  Med.  29,  409-480. 

1882  Edlefsen,  G.:  Ueber  die  Stoffwechselvorgange  beim   Fieber,   Mitt,   des 

Vereins  schleswig-holst.  Aerzte  1882,  No.  3;  abs.  Centralbl.  med.  Wis- 
sensch. 20  (1882),  783. 

1901  Edlefsen,  G.:  Ueber  die  Hauptunterschiede  zwischen  der  Kuhmilch  und 
Frauenmilch  und  den  Werth  und  die  Bedeutung  der  Ersatzmittel  fur 
Muttermilch,  Munch,  med.  Wochensch.  48,  7-11. 

1896  Edmunds,  Arthur:  Notes  on  Rennet  and  on  the  Coagulation  of  Milk, 
Journal  of  Physiology  19,  466-476. 

1903  Edsall  and  Miller:  A  Contribution  to  the  Chemical  Pathology  of  Acro- 
megaly, Med.  Bui.,  Univ.  Penn.  16,  143-150. 

1860  Edwards,  A,  M.:  Etudes  chimiques  et  physiologiques  sur  les  os,  Annales 
sci.  naturelles,  [4]  Zool.  14,  113-192. 

1900  Ehrlich,  Ernst:  Stoffwechselversuche  mit  P-haltigen  und  P-freien 
Eiweisskorpern,  Inaug.  Diss.,  Breslau.  28  pp. 

1913  Ehrmann,  R.,  and  E.  Kruspe:  Die  Verdauung  des  Lecithins  bei  Erkrank- 

ungen  des  Magendarmkanals,  Berlin  klin.  Wochenschr.  50,  1111. 
1903a  Ehrstrom,  Robert :  Zur  Kenntniss  des  Phosphorumsatzes  bei  dem  erwach- 
senen  Menschen,  Skand.  Arch.  Physiol.  14,  82-111. 


614  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1903b  Ehrstrom,  Robert:  Ueber  den  Nahrwerth  des  Caseinklystiere  nebst 
Bemerkimgen  iiber  den  Phosphorstoffwechsel,  Zeit.  klin.  Med.  49,  377- 
392. 

1903  Eichelberg,  Simon:  Ueber  den  Einfiuss  der  Driisengifte  Atropin  und 
Pilokarpin  auf  den  Stoffwechsel,  insbesondere  auf  die  Ausscheidung 
von  Stickstoff,  Phosphorsaure  und  Harnsaure,  Inaug.  Diss.,  Marburg. 
48  pp. 

1911  Eijkmann,  C:  Polyneuritis  Gallinarum  und  Beriberi,  Arch.  Schiffs-  u. 
Tropenhygiene  15,  698-712. 

1899  Ellenberger:  Die  Eigenschaften  der  Eselinmilch,  Arch.  Anat.  u.  Physiol. 
33-52. 

1902  Ellenberger:  Die  Zusammensetzung  und  die  Eigenschaften  der  Esel- 
inmilch, Ibid.  Suppl.,  313-322. 

1902  Ellenberger,  Seeliger  and  Klimmer:  Die  Eigenschaften  und  Zusammen- 

setzung der  Eselinmilch,  Arch,  wissensch.  u.  prakt.  Thierheilk.  28,  3 
and  4,  247-298;  through  Jahresb.  u.  d.  Fortschr.  d.  Thierchem.  32 
(1902),  989,  990. 
1895  Emery,  F.  E.,  and  B.  W.  Kilgore:  Fertilizing  Constituents  of  Various 
Rations  of  Cotton-seed  Hulls  and  Meal  Recovered  in  Manure  of 
Animals,  North  Carolina  Agr.  Exp.  Sta.  Bui.  118,  245-253. 

1909  Emmett,  A.  D.:  Chemistry  of  Animal  Feces.       Second  paper.       The 

Determination  of  Fatty  Matter  in  Animal  Feces  by  Ether  and  Carbon 
Tetrachloride,  Jour.  Amer.   Chem.   Soc.   31,   693-695;   871. 

1906  Emmett,  A.  D.,  and  H.  S.  Grindley:  The  Chemistry  of  Flesh.    III.      A 

Study  of  the  Phosphorus  Content  of  Flesh,  Ibid.  28,  25-63. 

1909a  Emmett,  A.  D.,  and  H.  S.  Grindley:  The  Chemistry  of  Flesh.  (Seventh 
paper.)  A  Preliminary  Study  of  the  Effect  of  Cold  Storage  upon  Beef 
and  Poultry.  (First  communication.)  Jour.  Ind.  and  Engin.  Chem. 
1,  413-436. 

1909b  Emmett,  A.  D.,  and  H.  S.  Grindley:  Chemistry  of  Flesh.  (Eighth  paper.) 
A  Preliminary  Study  of  the  Effect  of  Cold  Storage  upon  Beef  and 
Poultry,  (Second  communication.)       Ibid.  1,  580-597. 

1903  Enea,  Domenico:  Azione  della  nucleina  da    batterii    patogeni    e    non 

patogeni  sul  potere  battericida    del    siero    di    sangue    normale,    La 
Riforma  medica,  Napoli,  19,  1292-1296. 

1910  Engel,  and  A.  Frehn:  Der  Caseingehalt  der  Frauenmilch,  Berlin,  klin. 

Wochenschr.  47,  I,  436-438. 
1871     Engelmann,  Geo.  J.:   Schwefelsaure-  und  Phosphorsaure- Ausscheidung 

bei  korperliche  Arbeit,  Arch.  Anat.  u.  Physiol.  (1871)  14-30. 
1913     Eppler,  Julius:  Untersuchungen  iiber  Phosphatide,  insbesondere  iiber  die 

im  Eigelb  vorhandenen,  Zeit.  physiol.  Chem.  87,  233-254. 

1902  Erben,  Franz:  Ueber  die  chemische  Zusammensetzung  des  chlorotischen 

Blutes,  Zeit.  klin.  Med.  47,  302-320. 

1903  Erben,  Franz:  Studien  iiber  Nephritis.      I.    Ibid.  50,  441-463. 
1905     Erben,  Franz:  Studien  iiber  Nephritis.     II.    Ibid.  57,  39-69. 

1907  Erben,  Franz:  Ueber  den  Lezithingehalt  der  Erythrocyten  bei  Diabetes 

mellitus,  Zentralbl.  innere  Med.  28,  1090-1093. 

1908  Erben,  Franz :  Die  chemische  Zusammensetzung  des  leukamischen  Blutes, 

Zeit.  klin.  Med.  66,  278-301. 


PHOSPHORUS  METABOLISM  615 

1911  Erdheim,  J.:  Ueber  den  Kalkgehalt  des  wachsenden  Knochens  und  des 

Callus  nach  der  Epithelkorperchenexstirpation,  Frankfurter  Zeit. 
Path.  7,  175-230. 

1906  Erlandsen,  A.  W.  E.:  Undersogelser  over  Hjertets  Phosphatides       (Die 

Phosphatide  des  Herzens.)  Diss.,  Copenhagen.  157  pp.  Through 
Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  36  (1906),  495-498. 

1907  Erlandsen,  A.  W.  E.:  Untersuchungen  uber  die  lecithinartigen  Substan- 

zen  des  Myocardiums  und  der  quergestreiften  Muskeln,  Zeit.  physiol. 
Chem.  51,  71-155. 

1908  Errani,  Galdino:  Contributo  clinico  all'  uso  della  lecitina,   La'Rassegna 

di  terapia  3,  833-846.      75  refs. 
1874     Etzinger,  Johann:  Ueber  die  Verdaulichkeit  der  leimgebenden  Gewebe, 

Zeit.  Biol.  10,  84-110. 
1907    Euler,  Hans:  Gleichgewicht  und  Endzustand  bei  Enzymreaktionen,  Zeit. 

physiol.  Chem.  52,  146-158. 
1912a  Euler,  Hans:  Ueber  die  Wirkungsweise  der  Phosphatese.       III.  Mitteil- 

ung,  Biochem.  Zeit.  41,  215-223. 
1912b  Euler,  Hans:  Verhalten  der  Kohlenhydratphosphorsaureester  im  Tier- 

korper,  Zeit.  physiol.  Chem.  79,  375-397. 
1913     Euler,  Hans:  Om  Kolhydratfosforsyra  och     fosfaters     biokemiska     roll 

(Biochemical  Role  of  Phosphates  and  Carbohydrate-phosphoric  Acid 

Esters),  Svensk.  kern.  Tidskrift  25,  168-175;  through  Chem.  Abs.  8 

(1914),  715. 

1912  Euler,  Hans,  and  Helmer  Backstrom:  Zur  Kenntnis  der  Hefegarung,  II. 

Mitteilung,  Zeit.  physiol.  Chem.  77,  394-401. 
(1911)  Euler,  H.,  and  A.  Fodor:  Ueber  ein  Zwischenprodukt  der  alkoholischen 
Garung,  Biochem.  Zeit.  36,  401-410;  through  Jahresb.  ii.  d.  Fortschr.  d. 
Thierchem.  41  (1911),  782. 

1912  Euler,  Hans,  and  Yngoe  Funke:  Ueber  die  Spaltung  der  Kohlenhydrat- 

phosphorsaureester, Zeit.  physiol.  Chem.  77,  488-496. 
1912a  Euler,  Hans,  and  David  Johansson:  Ueber  den  Einflusz  des  Toluols  auf 

die  Zymasen  und  auf  die  Phosphatese,  Ibid.  80,  175-181. 
1912b  Euler,  Hans,  and  David  Johansson:  Versuche     uber     die     enzymatische 

Phosphatbindung,  Ibid.  80,  205-211. 

1913  Euler,. Hans,  and  David  Johansson:  Ueber  die  Reaktionsphasen  der  alko- 

holischen Garung,  Arkiv  Kemi,  Min.  Geol.,  4,  1-17;  through  Chem.  Abs. 
7  (1913),  3511. 

1911a  Euler,  Hans,  and  Sixten  Kullberg:  Untersuchungen  uber  die  chemische- 
Zusammensetzung  und  Bildung  der  Enzyme.  III.  Mitteilung.  Zeit. 
physiol.  Chem.  71,  14-30. 

1911b  Euler,  Hans,  and  Sixten  Kullberg:  Ueber  die  Wirkungsweise  der  Phos- 
phatese.     I.  Mitteilung.      Ibid.  74,  15-28. 

1911  Euler,  Hans,  and  Gunnar  Lundeqvist:  Zur  Kenntnis  der  Hefegarung, 
Ibid.  72,  97-112. 

1911  Euler,  Hans,  and  Hjalmar  Ohlsen:  Ueber  den  Einfluss  der  Temperatur 

auf  die  Wirkung  der  Phosphatese,  Biochem.  Zeit.  37,  313-320. 

1912  Euler,  Hans,  and  Hjalmar  Ohlsen:  Ueber  die  Wirkungsweise  der  Phos- 

phatese.     II.  Mitt.  Zeit.  physiol.  Chem.  76,  468-477. 
1911     Euler,  Hans,  and  Beth  af  Ugglas:  Untersuchungen  uber  die  chemische 
Zusammensetzung  und  Bildung  der  Enzyme.      II.    Mitteilung.      Ibid. 
70,  279-290. 


616  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1883     Ewald,  C.  A.:  Ueber  Phosphorsaureausscheidung  bei  Paralysis  agitans 

und  verwandten  Formen  der  Zitterlahmung,  Berlin,  klin.  Wochenschr. 

484-486;  502-504. 
1908    Ewing,  James:  Acidosis  and  Associated  Conditions,  Archives  of  Inter- 
nal Medicine  2,  330-354. 
1903     Fabiani,  Pietro:  II  potere  curativo  dei  glicerofosfati,  del  Cav.  Cesare 

Sebastiani,  Nuova  scuola  med.  napolitana  19,  fasc.  12,  1-8. 
1850     Falck,  C:  Die  Knochencaries  u.  ihr  Verhaltniss  zum  phosphors.  Kalk, 

Schmidt's  Jahrbiicher  der  in-  und  auslandischen  gesammten  Med.  67, 

261-263. 
1872    Falck,  C.  Ph. :  Ueber  die  Ausscheidung  von  in  das  Blut  gebrachtem  phos- 

phorsaurem  Natron  durch  die  Nieren,  Arch.  path.  Anat.  u.  Physiol.  54, 

173-184. 
1875     Falck,  F.  A.:  Ueber  den  Stoffwechsel  des  hungernden  Hundes,  Sitzungs- 

ber.  der  Gesell.  zur  Beforderung  der  ges.  Naturwissensch.  zu  Marburg, 

1875,  No.  7,  83-90. 
1908    Falk,  Fritz:  Ueber  die  chemische    Zusammensetzung    der    peripheren 

Nerven,  Biochem.  Zeit.  13,  153-172. 

1906  Falta,  W.,  and  C.  T.  Noeggerath:  Futterungsversuche  mit  kiinstlicher 

Nahrung,  Beitrage  z.  chem.  Physiol,  u.  Path.  7,  313-322. 
1908     Falta,  W.,  and  James  Lyman  Whitney:  Zur  Kenntnis  des  Eiweiss-  und 

Mineralstoffwechsels  pankreasdiabetischer  Hunde,  Ibid.  11,  224-228. 
1912     Fargier:  Beriberi  et  riz  fraichement  decortique,  Ann.  hyg.  med.  colon. 

15,  491-497. 

1907  Fauvel,  Pierre:  Quelques  experiences  sur     la    valeur     alimentaire    de 

differents  pains,  Revue  internat.  des  falsifications  20,  19,  20. 
1881     Feder,  Ludwig:  Der  zeitliche  Ablauf  der  Zersetzung  im  Thierkorper, 

Zeit.  Biol.  17,  531-576. 
1891     Fehling,  H.:  Ueber  Wesen  und  Behandlung    der    puerperalen     Osteo- 

malakie,  Arch.  Gynakol.  39,  171-196. 
1895     Fehling,  H.:  Weitere  Beitrage  zur  Lehre  von  der  Osteomalakie,  Ibid. 

48,  472^498. 

1912  Feinschmidt,  J. :  Die  Saureflockung  von  Lecithinen  und  Lecithin-Eiweiss- 

gemischen,  Biochem.  Zeit.  38,  244-251;     through     Zeit.     Unters.     d. 
Nahrungs-  u.  Genussmittel  24  (1912),  629. 

1913  Fenger,  Frederic:  On  the  Iodine  and    Phosphorus    Contents,    Size    and 

Physiological  Activity  of  the  Fetal  Thyroid  Gland,  Jour.  Biol.  Chem. 
14,  397-405. 

1914  Fenger,  Frederic:  The  Influence  of  Pregnancy  and  Castration  on  the 

Iodine  and  Phosphorus  Metabolism  of  the  Thyroid  Gland,  Ibid.  17, 

23-28. 
(1911)  Fernbach  and  Schoen:    (Einige  Bemerkungen     iiber     den     Reaktions- 

mechanismus  der  proteolytischen  Diastasen),  Compt.  rend.  Acad,  des 

sci.  153,  133-136;  through  Jahresb.  u.  d.  Fortschr.  d.  Thierchem.  41 

(1911),  732. 
1907    Ferrata,  A.,  and  G.  Moruzzi:  Ueber  das  Verhalten  von  Phosphorverbind- 

ungen    in    der     Darmschleimhaut    im     Hungerzustand     sowie     nach 

Verabreichung    von    Nahrungsstoffen,    Arch.    Verdauungskrankh.    13, 

223-232. 


PHOSPHORUS  METABOLISM  617 

1910  Fetzer,  L.  W.:  The  Lecithin  Content  of  Milk  under  Pathologic  Conditions, 
Meeting  Amer.  Assoc,  for  the  Adv.  of  Sci.,  1910,  report  in  Science, 
n.  s.  33  (1911),  339. 

1912  Feulgen,  R.:  Das  Verhalten  der  echten  Nucleinsaure  zu  Farbstoffen.  1. 
Mitteilung,  Zeit.  physiol.  Chem.  80,  73-78. 

1913a  Feulgen,  R.:  Das  Verhalten  der  echten  Nucleinsaure  zu  Farbstoffen.  II 
Mitteilung,  Ibid.  84,  309-328. 

1913b  Feulgen,  R.:  Ueber  eine  Nucleinsaure  aus  der  Pankreasdriise.  I.  Mit- 
teilung, Ibid.  88,  370-376. 

1911a  Fingerling,  Gustav:  Die  Verwertung  der  in  den  Futtermitteln  enthalten- 
en  Phosphorverbindungen  durch  Wiederkauer,  Biochem.  Zeit.  37,  266- 

271. 

1911b  Fingerling,  Gustav:  Beitrage  zur  Verwertung  von  Kalk-  und  Phosphor- 
saureverbindungen  durch  den  tierischen  Organismus.  I.  Einfluss 
kalk-  und  phosphorsaurearmer  Nahrung  auf  die  Milchsekretion,  Land- 
wirtsch.  Versuch.  Stat.  75,  1-52. 

1912a  Fingerling,  Gustav:  Die  Bildung  von  organischen  Phosphorverbindungen 
aus  anorganischen  Phosphaten,  Biochem.  Zeit.  38,  448-467. 

1912b  Fingerling,  Gustav:  Einflusz  organischer  und  anorganischer  Phosphor- 
verbindungen auf  die  Milchsekretion,  Ibid.  39,  239-268. 

(1913)  Fingerling,  Gustav:  (The  Utilization  of  Calcium  and  Phosphorus  Com- 
pounds by  the  Animal  Organism.  II.  The  Use  of  the  More  Important 
Phosphorus  Compounds  by  Ruminants),  Landwirtsch.  Versuch.  Stat. 
79-80,  847-870;  through  Chem.  Abs.  7  (1913),  3355. 

1903  Finzi,  Roberto:  Modificazioni  della  composizione  della  saliva  per  alter- 
azioni  della  crasi  del  sangue  dovute  ad  iniezioni  endovenosa  di  fosfato 
neutro  di  sodio,  Bollett.  delle  sci.  med.  di  Bologna  74,  23-32. 

1903  Fischer,  Emil,  and  Emil  Abderhalden:  Ueber  die  Verdauung  des  Caseins 
durch  Pepsinsalzsaure  und  Pankreasfermente,  Zeit.  physiol.  Chem.  40, 
215-219. 

1894  Fischer,  Heinrich:  Casuistischer  Beitrag  zur  Behandlung  der  Osteomal- 
acie,  Prager  med.  Wochenschr.  19,  425-427. 

1907  Fitz,  R.,  C.  L.  Alsberg  and  L.  J.  Henderson:  Concerning  the  Excretion  of 
Phosphoric  Acid  during  Experimental  Acidosis  in  Rabbits,  Amer.  Jour. 
Physiol.  18,  113-122. 

1907  Fjodoroff,  Vladimir  Stepanovitch :  Ueber  den  Einfluss  organischer  Phos- 
phorpraparate  (Lecithin,  Nucleinsaure,  Phytin  und  Protylin)  auf  die 
Magenverdauung,  Diss.,  St.  Petersburg,  103  pp.  Through  Russische 
med.  Rundschau  6  (1908),  729-731. 

1907  Flamini,  M.:  L'azione  del  fosforo  sul  ricambio  del  calcio  in  bambini 
normali  ed  in  bambini  rachitici,  Archivio  di  farmacol.  6,  653-663. 

1881  Fleischer,  R.:  Klinische  und  pathologisch-chemische  Beitrage  zur  Lehre 
von  den  Nierenkrankheiten,  Deut.  Arch.  klin.  Med.  29,  129-192. 

1906  Fleischner,  E.  C:  The  Relation  of  Weight  to  the  Measurements  of  Child- 
ren During  the  First  Year,  Archives  of  Pediat.  23,  739-757. 

1876  Flesch,  M.:  Zur  Physiologie  der  Knochen-Resorption,  Centralbl.  med. 
Wissensch.  14,  524,  525. 

1909  Fletcher,  William:  Rice  and  Beriberi,  Jour,  of  Tropical  Med.  and  Hyg.  12, 
127-135. 

1897  Flieger,  Erdmann:  Ueber  die  Behandlung  der  Rhachitis  mit  Phosphor, 
Inaug.  Diss.,  Breslau.       39  pp. 


618  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1871  Flint,  Austin,  Jr.:  On  the  Physiological  Effects  of  Severe  and  Protracted 
Muscular  Exercise;  with  Especial  Reference  to  the  Influence  of  Exer- 
cise upon  the  Excretion  of  Nitrogen,  N.  Y.  Med.  Jour.  13,  609-697. 

1900  Forberg:  Welchen  Einfluss  iiben  Nahrungsmittel  und  Trinkwasser  auf 
Entwicklung  und  Bestand  der  Zahne  aus  ?  Odontolog.  Blatter  1900, 
Nr.  24;  through  Centralbl.  Stoffwechsel  u.  Verdauungskrankh.  II 
(1901),  107. 

1873  Fokker,  A.  P. :  Ueber  das  Vorkommen  von  gelosten  Erden  und  Phosphor- 
saure  im  alkalischen  Blute,  Arch.  ges.  Physiol.  7,  274-284. 

1905  Folin,  Otto:  Laws  Governing  the  Chemical  Composition  of  Urine,  Amer. 
Jour.  Physiol.  13,  66-115. 

1907  Folin,  Otto:  The  Acid  Intoxication  Theory,  Jour.  Amer.  Med. 
Assoc.  49,  128-131. 

1912  Folin,  Otto,  and  W.  Denis :  Tyrosine  in  Proteins  as  Determined  by  a  New 

Colorimetric  Method,  Jour.  Biol.  Chem.  12,  245-251. 
1902     Folin,  Otto,  and  Philip  Shaffer:  On  Phosphate  Metabolism,  Amer.  Jour. 

Physiol.  7,  135-151. 
1904    Folin,  Otto,  Philip  A.  Shaffer,  and  L.  A.  Hill:  Some  Metabolism  Studies 

with  Special  Reference  to  Mental  Disorders,  Amer.  Jour.  Insanity  60, 

699-732;  61,  299-364. 

1909  Forbes,  E.  B.:  Specific  Effects  of  Rations  on  the  Development  of  Swine, 

Ohio  Agr.  Exp.  Sta.  Bui.  213,  239-305;  Mo.  Agr.  Exp.  Sta.  Bui.  81,  3-69. 
1914    Forbes,  E.  B.,  and  associates:  The  Metabolism  of  Organic  and  Inorganic 

Phosphorus  Compounds,  Ohio  Agr.  Exp.  Sta.  Research  Bui.  6. 
1914    Forbes,  E.  B.,  F.  M.  Beegle,  C.  M.  Fritz  and  J.  E.  Mensching:  A  Chemical 

Study  of  the  Nutrition  of  Swine,  Ohio  Agr.  Exp.  Sta.  Bui.  271,  225-261. 

1913  Forbes,  E.  B.,  F.  M.  Beegle,  and  J.  E.  Mensching:  Mineral  and  Organic 

Analyses  of  Foods,  Ohio  Agr.  Exp.  Sta.  Bui.  255,  211-231. 

1914  Forbes,  E.  B.,  and  C.  M.  Fritz:  The  Effects  of  the  Ensilage  Process  on 

the  Solubility  of  Floats,  Jour.  Ind.  and  Engin.  Chem.  6,  222,  223. 

1910  Forbes,  E.  B.,  A.  Lehmann,  R.  C.  Collison,  and  A.  C.  Whittier:  Methods 

for  the  Quantitative  Estimation  of  Inorganic  Phosphorus  in  Vegetable 

and  Animal  Substances,  Ohio  Agr.  Exp.  Sta.  Bui.  215,  459-489. 
1910     Forbes,  E.  B.,  A.  C.  Whittier,  and  R.  C.  Collison:  The  Mineral  Nutrients 

of  Blue-Grass,  Ohio  Agr.  Exp.  Sta.  Bui  222,  39-52. 
1873a  Forster,  J.:  Versuche  liber  die  Bedeutung  der  Aschenbestandteile  in  der 

Nahrung,  Zeit.  Biol.  9,  297-380. 
1873b  Forster,  J.:  Beitrage  zur  Ernahrungsfrage,  Ibid.  9,  381-410. 
1876     Forster,  J.:  Ueber  die  Verarmung  des  Korpers,  speciell  der  Knochen  an 

Kalk  bei  ungeniigender  Kalkzufuhr,  Ibid.  12,  464-474. 
1884    Forster,  J.:  Beitrage  zur  Kenntnis  der  Kalkresorption  im  Thierkorper, 

Arch.  Hyg.  2,  385-411. 
1793    Fourcroy,  A.  F.:  Examen  chimique  du  cerveau  de    plusieurs    animaux, 

Annales  de  chim.  16,  282-322. 
1908    Frankel,  Sigmund:  Ueber  Lipoide,  Verhandl.  des  Kongr.  innere  Med.  25, 

564-569. 
1909a  Frankel,  Sigmund:  Gehirn-Chemie,  Ergebnisse  d.  Physiol.  8,  212-253.  75 

refs. 


PHOSPHORUS  METABOLISM  619 

1909b  Frankel,  Sigmund:  Ueber  Lipoide.  VI.  Mitteilung.  Ueber  eines  neues 
Verfahren  der  fraktionierten  Extraktion  der  Gehirnlipoide,  Biochem. 
Zeit.  19,  254-265. 

1908  Frankel,   Sigmund,  and  Carlo  Bolaffio:   Ueber  Lipoide.     I.   Ueber  das 

Neottin,  ein  Triaminomonophosphatid,  Ibid.  9,  44-53. 

1909  Frankel,   Sigmund,   and   Ludwig   Dimitz:   Ueber  Lipoide.       VIII.  Mitt. 

Ueber  die  Spaltungsprodukte  des  Kephalins,  Ibid.  21,  337-347. 

1910  Frankel,  Sigmund,  (and  Kurt  Linnert) :  Ueber  Lipoide.      IX.  Mitt.  Ueber 

das  Sahidin  aus  Menschenhirn,  Ibid.  24,  268-276. 
1909     Frankel,  Sigmund,  Kurt  Linnert  and  G.  A.  Pari:  Ueber  Lipoide.    V.  Mitt. 

Ueber  die  Phosphatide  des  Rinderpankreas,  Ibid.  18,  37-39. 
1909     Frankel,  Sigmund,  and  Ernst  Neubauer:  Ueber     Lipoide,      VII.  Mitt. 

Ueber  Kephalin,  Ibid.  21,  321-336. 
1909a  Frankel,  Sigmund,  and  Alexander  Nogueira:  Ueber  Lipoide.       II.  Mitt. 

Ueber  die  ungesattigten  Phosphatide  der  Niere,  Ibid.  16,  366-377. 
1909b  Frankel,  Sigmund,  and  Alexander  Nogueira:  Ueber  Lipoide.      III.  Mitt. 

Ueber  die  Wechselwirkung  der  ungesattigten  Nierenphosphatide  mit 

Farbstoffen,  Ibid.  16,  378-382. 

1909  Frankel,  Sigmund,  and  G.  A.  Pari:  Ueber  Lipoide.      IV.  Mitt.     Ueber 

die  Phosphatide  des  Rinderpankreas,  Ibid.  17,  68-77. 
1904     Franchini,  Giuseppe:  Sul  ricambio  materiale  in  uno  caso  di  acromegalia, 

Bollett.  delle  sci.  med.  di  Bologna  75  (4  of  ser.  8),  8;  through  Biochem. 

Centralbl.  3  (1905),  522. 
1907     Franchini,  Giuseppe:  Ueber  den  Ansatz  von  Lecithin  und  sein  Verhalten 

im  Organismus,  Biochem.  Zeit.  6,  210-225. 
1908a  Franchini,  Giuseppe:  Ricerche  sulla  lecitina,  colina  et  acido  formico, 

Archivio  di  farmacologia  7,  371-389. 
1908b  Franchini,  Giuseppe:  Beitrag  zum  chemischen  und  histologischen  Studium 

des  Blutes  bei  Akromegalie,  Berlin,  klin.  Wochenschr.  45,  II,  1636-1639. 

1910  Franchini,  Giuseppe:  Die  Funktion  der  Hypophyse  und  die  Wirkungen 

der  Injektion  ihres  Extractes  bei  Tieren,  Ibid.  47,  I,  613-617;  670-673. 
719-723. 

1910  Francis,  C.  K.,  and  P.  F.  Trowbridge:  Phosphorus  in  Beef  Animals.  Parts 
I  and  II,  Jour.  Biol.  Chem.  7,  481-501;  8,  81-93. 

1913  Francois,  M.,  and  E.  Boismenu:  Le  glycerophosphate  de  calcium  commer- 
cial, Jour,  de  pharm.  et  de  chim.  [ser.  7]  7,  448-457,  492-497. 

1909  Frank,  Franz,  and  Alfred  Schittenhelm:  Ueber  die  Umsetzung  verfiit- 

terte  Nucleinsaure  beim  normalen  Menschen,  Zeit.  physiol.  Chem.  63, 
269-282. 

1910  Frank,  L. :  Ueber  den  Einfluss  kalkarmen  Futters  auf  den  kalkgehalt  der 

Kuhmilch,  Chem.  Zeitung  34,  978,  979. 
1894    Frankfurt,  Saloman:  Ueber  die  Zusammensetzung  der  Samen  und  der 

etiolierten  Keimpflanzen  von  Cannabis  sativa  und  Helianthus  annuus, 

Landwirtsch.  Versuch.  Stat.  43,  143-179. 
1912    Fraps,  G.  S.,  and  J.  B.  Rather:  Composition  and  Digestibility  of  the 

Ether  Etract  of  Hays  and  Fodders,  Texas  Agr.  Exp.  Sta.,  Tech.  Bui. 

No.  150.      29  pp. 
1910    Fraser,  Henry,  and  A.  T.  Stanton:  The  Etiology  of  Beriberi.  Philippine 

Jour,  of  Sci.,  B,  5,  55-64. 


620  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1911a  Fraser,  Henry,  and  A.  T.  Stanton:  The  Etiology  of  Beriberi,  Studies 

Inst.  Med.  Research,  Federated  Malay  States,  No.  12;  through  Jour. 

of  Tropical  Med.  and  Hyg.  14  (1911),  248,  249. 
1911b  Fraser,  Henry,  and  A.  T.  Stanton:  The  Etiology  of  Beriberi,  Jour,  of 

Tropical  Med.  and  Hyg.  14,  333-341;  349-358;  365-374. 
1914    Fraser,  Henry,  and  A.  T.  Stanton:  Unpolished  Rice  and  the  Prevention 

of  Beriberi,  The  Lancet  186,  96-98. 

1904  Fratta,  Aldo:  Rapporto  fra  l'azione  della  cura  antirabica  e  quella  del 

glicerofosfati  e  della  lecitine,  Bollett.  delle  sci.  med.  di  Bologna  75, 
593-610. 

1841  Fremy,  Edmond :  Recherches  sur  le  cerveau,  Annales  de  chim.  et  de  phys. 

173  (2  of  ser.  3);  463-488;  also  Jour,  de  Pharm.  27,  453. 

1842  Fremy,  Edmond:  Ueber  das  Gehirn,  Jour.  f.  prakt.  Chem.  25,  29-50. 
1855     Fremy,  Edmond :  Recherches  chimiques  sur  les  os,  Annales  de  chim.  et  de 

phys.  214  (43  of  ser.  3),  47-109. 
1842    Frerichs:  Ueber  die    chemische    Zusammensetzung    der    menschlichen 

Knochen,  Annalen  der  -Chem.  u.  Pharm.  43,  251-255. 
1891     Freund,  E.,  and  F.  Obermayer:  Ueber  die  chemische  Zusammensetzung 

leukamischen  Blutes,  Zeit.  physiol.  Chem.  15,  310-318. 
1901     Freund,  E.  and  O.:  Beitrage  zum  Stoffwechsel  im  Hungerzustand,  Wien. 

klin.  Rundschau  15,  69-91. 

1905  Freund,  W.:  Zur  Wirkung  der  Fettdarreichung  auf  den  Sauglingsstoff- 

wechsel,  Jahrb.  Kinderheilk,  61,  36-50. 

1906  Frey,   Ernst:   Therapeutischer  Wert  der   Glycerophosphate,   Budapest! 

Orvosi  Ujsag  4,  145;  through  Pester  med.-chir.  Presse  42  (1906),  862, 
1911     Frey,  Ernst:  Jodid,  Nitrat,  Sulfat,  Phosphat  werden  durch  Sekretion  in 

den  Harnkanalchen  ausgeschieden,  Arch.  ges.  Physiol.  139,  512-531. 
1911     Friedenthal,  Hans:  Ueber  die  massgebende  Rolle  der  Salze  der  Frauen- 

milch  bei  der  Ernahrung  im  Sauglings-  und  ersten  Kindesalter,  Munch. 

med.  Wochenschr.  58,  II,  2385-2387. 

1911  Fromherz,  K.:  Zur  Kenntniss  der  Wirkungsweise  der  Phenylcinchonin- 

saure  auf  den  Purinstoffwechsel  des  Hundes,  Biochem.  Zeit.  35,  494- 
502. 

1912  Frouin,  Albert,  and  Pierre  Gerard:  Sur  la  composition  minerale  du  sue 

pancreatique  de  chien  et  de  vache,  Compt.  rend.  Soc.  de  Biol.  72,  98-100. 

1903  Fiirst,   L. :  Ueber  Ernahrungs-  und  Funktionsinsuffizienz  des   Gehirns, 

Therapeut.  Monatsh.  16,  No.  8;  through  Jour.  Amer.  Med.  Assoc.  41, 
Pt.  II,  1378. 

1904  Fiirst :  Einige  Erf ahrungen  iiber  des  Phytin  als  Antirhachiticum  und  als 

Mittel  gegen  Psychasthenie,  Centralbl.  Kinderheilk.      9,  409-413. 

1907  von  Fiirth,  Otto,  and  Ernst  Jerusalem:  Ueber  die  chemische  Stellung  der 

Pankreasnucleinsaure   (Guanylsaure),  Beitrage  z.  chem.  Physiol,  u. 
Path.  10,  174-187. 

1908  von  Fiirth,  Otto,  and  Ernst  Jerusalem:     Ueber  die  chemische  Stellung 

der  Pankreasnucleinsaure  (Guanylsaure),  II.  Mitteilung,  Ibid.  11,  146- 
150. 
1910    Fujitani,  I.:  Contribution  to  the  Etiology  of  the  Disease  Resulting  from 
Feeding  Birds  on  Rice,  Archives  internat.  de  pharmacodyn.  et  de  ther# 
20,  288-309. 


PHOSPHORUS  METABOLISM  621 

1902a  Fuld,  Ernst:  Ueber  die  Verbindungen  von  Eiweisskorpern  mit  Metaphos- 

phorsaure,  Beitrage  z.  chem.  Physiol,  u.  Path.  2,  155-168. 
1902b  Fuld,  Ernst :  Ueber  Milchgerinnung  durch  Lab,  Ergebnisse  d.  Physiologie 

1,  Abt.  1,  468-504.      114  refs. 
1907    Funatsu,  T.:  On  Different  Forms  of  Phosphoric  Acid  in  Press  Cakes, 

Bui.  Coll.  Agr.  Tokyo  7,  457-459. 
1911     Funk,  Casimir:  The  Chemical  Nature  of  the  Substance  which  Cures 

Polyneuritis  in  Birds  Induced  by  a  Diet  of  Polished  Rice,  Jour,  of 

Physiol.  43,  395-400. 
1912a  Funk,  Casimir:  The  Effect  of  a  Diet  of  Polished  Rice  on  the  Nitrogen 

and  Phosphorus  of  the  Brain,  Ibid.  44,  50-53. 
1912b  Funk,  Casimir:  Preparation  from  Yeast  and  Certain  Foodstuffs  of  the 

Substance  the  Deficiency  of  Which  in  Diet  Occasions  Polyneuritis  in 

Birds,  Ibid.  45,  75-81. 
1912c  Funk,  Casimir:  The  Substance  from  Yeast  and  Certain  Foodstuffs  which 

Prevents  Polyneuritis (  Beriberi),  British  Med."  Jour.,  1912,  II,  787,  788. 
1913     Funk,   Casimir:   Studies  on  Beriberi.       Further  Facts  Concerning  the 

Chemistry  of  the  Vitamine-fraction  from  Yeast,  Ibid.  1913,  I,  814. 
1894    Gabriel,  S.:  Chemische  Untersuchungen     fiber     die    Mineralstoffe     der 

Knochen  und  Zahne,  Zeit.  physiol.  Chem.  18,  257-302. 
1900    Galeotti,  Gino:  Azione  dei  nucleoproteidi  sulle  cellule     e     sui    tessuti. 

Ricerche  sperimentali,  Lo  Sperimentale  54,  435-463. 

1905  Galimard,  J.,  and  P.  Konig:  Recherches  sur  l'analyse  des  os  dans  un  cas 

d'osteomalacie  infantile,  Jour,  de  pharm.  et  de  chim.  21  (ser.  6),  352- 
357. 

1906  Gallenga,  Pietro:  Contributo  alio  studio  della  medicazione  fosforato  con 

i  preparati  organici  di  fosforo,  II  Policlinico  13,  115-129;  167-174. 
1879a  Gamgee,  Arthur,  and  Ernst  Blankenhorn:  Ueber  Protagon,  Arch.  path. 

Anat.  u.  Physiol.  77,  389-397. 
1879b  Gamgee,  Arthur,  and  Ernst  Blankenhorn.  Ueber  Protagon,  Ber.  deut. 

chem.  Gesell.  12,  1229-1234. 
1879c  Gamgee,  Arthur,  and  Ernst  Blankenhorn:  Ueber  Protagon,  Zeit.  physiol. 

Chem.  3,  260-283. 
1880     Gamgee,  Arthur,  and  Ernst  Blankenhorn:  Ueber     Protagon,     Zeit.    f. 

physiol.  Chem.  3,  260-283. 
1903a  Gamgee,  Arthur,  and  Walter  Jones:    Ueber    die    Nucleoproteide     des 

Pankreas,  der  Thymus  und  der  Nebenniere,  mit  besonderer  Berfick- 

sichtigung  ihrer  optischen  Aktivitat,  Beitrage  z.  chem.  Physiol,  u. 

Path.  4,  10-22. 
1903b  Gamgee,  Arthur,  and  Walter  Jones:  On  the  Nucleoproteids  of  the  Pan- 
creas, Thymus,  and  Suprarenal   Gland,  with  Especial  Reference  to 

their  Optical  Activity,  Amer.  Jour.  Physiol.  8,  447-455. 
1903c  Gamgee,  Arthur,  and  Walter  Jones:  Die  optische  Activitat  der  Nucleo- 

prote'ide  des  Pankreas,  des  Thymus  und  der  Nebennieren,  Ber.  deut. 

chem.  Gesell.  36,  914. 
1876-   Gamgee,  Arthur,  John  Priestley  and  Leopold  Larmuth:  On  the  Differ- 
1877    ence  in  the  Poisonous  Activity  of  Phosphorus  in  Ortho,-  Meta-  and 

Pyro-phosphoric  Acids,  Jour,  of  Anat.  and  Physiol.  11,  255-272. 
1898     Garratt,  G.  C:  On  the  Sequence  of  Certain     Changes     in     the     Urine 

Produced  by  Exercise  and  by  Turkish  Baths,  Jour,  of  Physiol.  23, 

150-162. 


622  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1904    Garrat,  G.  C:  Observations  on  Metabolism  in  the  Febrile  State  in  Man, 
Medico-Chirurg.  Transact.  87  (69  of  ser.  2),  163-326. 

1908  Gassmann,  Theodore:     Chemische     Untersuchungen     der     Zahne,     Zeit. 

physiol.  Chem.  55,  455-465. 

1910  Gassmann,  Theodore:  Chemische  Untersuchungen  von     gesunden     und 

rachitischen  Knochen,  Ibid.  70,  161-170. 
1913     Gassmann,     Theodore:     Die     Darstellung     eines     dem     Apatit-Typus 

entsprechenden  Komplexsalzes  und  seine  Beziehungen  zum  Knochenbau, 

Ibid.  83,  403-408. 
1902    Gaston,  Zephirin,  Octave,  Laurent-Boulenger :  Das  Lecithin,  durch  seine 

Anwendung  im  Organismus   erzeugte  Reaktionen.       These  de  Lille, 

1902.       136  pp.       Through  Jahresb.  u.  d.  Fortschr.  d.  Thierchem.  32 

(1902),  659. 
1895    Gaube,  J.:  La  mineralisation  du  lait,  Compt.  rend.  Soc.  de  biol.  47,  489, 

490. 

1909  Gaucher,  Louis:  Sur  la  digestion  gastrique  de  la"  caseine,  Compt.  rend. 

Acad,  des  sci.  148,  53-56;  also  Compt.  rend.  Soc.  de  biol.  66,  25-27;  536- 
538. 

1911  Gaucher,  Louis:     Sur     la     digestion     de     la     caseine,     Compt.     rend. 

Acad,  des  sci.  153,  891,  892. 

1912  Gaucher,  Louis :  Recherches  sur  la  digestion  du  lait.    Digestion  gastrique 

du  caseine,  Compt.  rend.  Soc.  de  biol.  72,  354,  355. 
1877-    Geoghegan,   Edward   G.:   Ueber   die   anorganischen   Gehirnsalze,   nebst 
1878    einer  Bestimmung  des  Nucleins  im  Gehirn,  Zeit.  physiol.   Chem.  1, 

330-338. 
1911     Gerard,  Ern.,  and  M.  Verhaeghe:  Contribution  a.  l'etude  chimique  des 

lipoi'des  des  organes  animaux,  Jour,  de  pharm.  et  de  chim.  3  (ser.  7), 

385-390. 
1875     Gerber,  N.:  Recherches  sur  le  lait,  Bui.  Soc.  chim.  28  (23  of  ser.  2),  342- 

344. 
1899     Gerhardt,  Dietrich,  and  Wilhelm     Schlesinger:    Ueber  die    Kalk-    und 

Magnesiaausscheidung  bei  Diabetes  mellitus  und  ihre  Beziehung  zur 

Ausscheidung  abnormer  Sauren  (Acidose),  Arch.  exp.  Path.  u.  Pharm. 

42,  83-108. 

1913  Gessner,  W.:  Ueber  die  Wirkung  des  Phosphors  im  Phosphorlebertran 

bei  Rachitis  als  Inflammator,  Berlin,  klin.  Wochenschr.  50,  688-690. 
1901     Gevaerts,  Jacques:  Diete  sous  phosphore,  La  Cellule  18,  7-33. 
1895     Giacosa,  G.:  Analyse  des  cendres  d'un  nouveau-ne,  Archives  ital.  de  biol. 

22,  252-258. 

1904  Giacosa,   Piero:   Sulla     fitina     (sale     calcico-magnesiaco     dell'     acido 

anidrossimetilendifosforico)     e     suo     comportarsi     nell*     organismo, 
Giornale  d.  reale  Accad.  di  med.  di  Torino  67,  414-416. 

1905  Giacosa,  Piero:  Sul  comportamento  nell'  organismo  della  fitina,  Ibid.  68, 

369-374. 
1905-    Giacosa,  Piero:  SulP  azione  della  fitina  in  relazione  alia  funzione  glico- 

1906     genica,  Atti  d.  reale  Accad.  sci.  di  Torino  41,  651-655. 
1907     Giacosa,  Piero:  SulP  azione  farmacologica  della  fitina,  Giornale  d.  reale 

Accad.  di  med.  di  Torino  70,  290-295. 
1909    Giacosa,  P.,  and  S.  Dezani :  Studi  sulla  secrezione  stomacale,  Atti  d.  reale 

Accad.  d.  sci.  di  Torino  44,  521-534. 


PHOSPHORUS  METABOLISM  623 

1905    Gianasso,  A.  B.,  and  V.  Ovazza:  Valore  terapeutico  della  fitina  in  pedia- 

tria,  Giornale  di  ginecol.  e  di  pediat.  5,  227-231. 
1899    Giertz,  K.H.:  Zur  Kenntniss  der  Pseudonucleine,  Zeit.  physiol.  Chem.  28, 

115-122. 
1907    Gies,  William  J.:  Further  Observations  on  Protagon,  Jour.  Biol.  Chem. 

3,  339-358. 
1913    Giffhorn,  Heinrich:  Beitrage  zur  Kenntnis  des  Stoffwechsels,  besonders 

der  Mineralien,  im  Sauglingsalter.  III.      Der  Einfluss  von  Fettzulagen 

auf  den  Stoffwechsel  verdauungsgesunder  Kinder  bei  molkenarmer  und 

molkenreicher  Ernahrung,  Jahrb.  f.  Kinderheilk.  78,  531-564. 
1901    Gilbert,  A.:  Lecithin  in  Therapeutics,  Therapeut.  Monatsh.,  Nov.  1901; 

through  Jour.  Amer.  Med.  Assoc.  37,  Pt.  II,  (1901),  1711. 
1901    Gilbert,  A.,  and  L.  Fournier:  La  lecithine  en  therapeutique,  Compt.  rend. 

Soc.  de  biol.  53, 145-148. 

1904  Gilbert,  A.,  and  A.    Lippmann:    Du    principe    phospho-organique    des 

graines  vegetales.        L'acide     anhydro-oxymethylene-diphosphorique, 
La  Presse  med.  545-547;  577-580. 
1903    Gilbert,  A.,  and  S.  Posternak:  La  medication  phosphoree  envisagee  au 
point  de  vue  des  echanges  nutritifs  de  l'organisme  (etude  critique  et 
experimental ),  L'Oeuvre  med.  -chir.,  No.  36,  48  pp.      117  refs. 

1905  Gilbert,  A.,  and  S.  Posternak:  Ueber  die  Phosphortherapie  vom  Stand- 

punkte  des  Stoffwechsels     aus     betrachtet,     Aertzliche     Rundschau 

(Munchen)  15,  398-402;  411-415. 
1888    Gilson,  Eugen:  Beitrage  zur  Kenntniss  des  Lecithins,  Zeit.  physiol.  Chem. 

12,  585-602. 
1884    Girard,  Aime:  Memoire  sur  la  composition  chimique  et  la  valeur  alimen- 

taire  des  diverses  parties  du  grain  de  froment,  Annales  de  chim.  et  de 

phys.  3  (ser.  6),  289-355. 
1903    Girard,  Aime,  and  L.  Lindet:  Le  froment  et  sa  mouture.       Traite  de 

meunerie,  355  pp. 
1907    Glikin,  W.:  Ueber  den  Lecithingehalt  des  Knochenmarks  bei  Tieren  und 

beim  Menschen,  Biochem.  Zeit.  4,  235-243. 
1908a  Glikin,  W.:  Zur  biologischen  Bedeutung  des  Lecithins.      I.  Mitteilung, 

Ibid.  7,  286-298. 
1908b  Glikin,  W.:  Ueber  den  Eisengehalt  der  Fette,  Lipoide  und  Wachsarten, 

Ber.  deut.  chem.  Gesell.  41,  910-915. 
1909a  Glikin,  W.:  Zur  biologischen  Bedeutung  des  Lecithins.      II.  Mitt.  Ueber 

den     Lecithingehalt  bei     Degenerationen     im     Zentralnervensystem, 

Biochem.  Zeit.  19,  270-273. 
1909b  Glikin,  W.:  Zur  biologischen  Bedeutung  des  Lecithins.      III.  Mitt.  Ueber 

den  Lecithin-  und  Eisengehalt  in  der  Kuh-  und  Frauenmilch,  Ibid.  21, 

348-354. 
1909c  Glikin,  W.:  Zur  biologischen  Bedeutung  des  Lecithins.      IV.  Mitt.  Ueber 

den  Gehalt  des  Blutes  bei  Polycythaemia  rubra  megalosplenica  an 

Phosphor  und  Eisen  in  Lipoidform,  Ibid.  22,  461-463. 
1903     Gnezda,  Julius:  Ueber  "Protylin,"  Deut.  Aerzte-Zeitung,   193-196. 
1844    Gobley,  Nicholas  Theodore:  De  la  presence  du  phosphore  dans  l'huile  de 

foie  de  raie,  Jour,  de  pharm.  et  de  chim.  6  (ser.  3),  25,  26. 
1846    Gobley,  Nicholas  Theodore:  Recherches  chimiques  sur  le  jaune  d'oeuf, 

Ibid.  9,  81-91;  161. 


624  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1847     Gobley,  Nicholas  Theodore:  Recherches  chimiques  sur  le  jaune  d'oeuf, 

deuxieme  memoire,  presente  a  l'Academie  royale  des  sciences.  Examen 

comparatif  du  jaune  d'oeuf  et  de  la  matiere  cerebrale  Ibid.  11  (ser.  3), 

409-417;  12   (ser.  3),  1-13. 
1850a  Gobley,  Nicholas  Theodore:  Recherches  chimiques  sur  les  oeufs  de  carpe, 

Ibid.  17  (ser.  3),  401-417;  18  (ser.  3),  107-119. 
1850b  Gobley,   Nicholas   Theodore:   Recherches   chimiques   sur  la   laitance   de 

carpe,  Ibid.  19  (ser.  3),  406-421. 
1851     Gobley,  Nicholas  Theodore:  Recherches  chimiques     sur     les     matieres 

grasses  du  sang  veineux  de  l'homme,  Ibid.  21  (ser.  3),  241-254. 
1856     Gobley,  Nicholas  Theodore:  Recherches  sur  la  nature  chimique  et  les 

proprietes  des  matieres  grasses  contenus  dans  la  bile,  Ibid.  30    (ser.  3), 

241-255. 
1877     Gobley,  Nicholas  Theodore:  Chemische  Untersuchungen  fiber  das  Gehirn, 

Arch.  Pharmacie  10  (ser.  3),  445-452. 

1903  Godefroy,  Maurice:  Recherches  sur  1 'elimination  du  phosphore  urinaire 

dans  les  cas  graves  de  rhumatisme  chronique,  These  de  Paris.     52  pp. 

1861  von  Gohren,  Theodor:  Ueber  eine  Beigabe  von  Knochenerde  zum  Futter 
der  Thiere,  nebst  einem  physiologisch-chemischen  Versuch  liber  deren 
Verdaulichkeit,  Landwirtsch.  Versuch.  Stat.  3,  161-178. 

1865  von  Gohren,  Theodor:  Knochen  von  einem  knochenbruchigen  Rinde, 
Centralbl.  ges.  Landescultur  in  Bohmen,  1865,  p.  344;  through 
Jahresb.  u.  d.  Fortschr.  d.  Agricultur-Chemie,  for  1866,  9  (1868),  345, 
346. 

1904  von  Gohren,  Theodor:  Schweinemilch,  Milch-Zeitung  33,  777. 

1907     Goldfarb,  A.  J.:  A  Study  of  the  Influence  of  Lecithin  on  Growth,  Proc. 

Soc.  Exp.  Biol,  and  Med.  4,  159,  160. 
1910     Goldfarb,  A.  J. :  Does  Lecithin  Influence  Growth  ?      Arch.  Entwicklungs- 

Mechanik  der  Organismen,  29,  255-274. 
1910     G61dingr  John,   and  S.  G.  Paine:   Note  upon  the  Composition   of  Milk 

Yielded   by    Cows    Fed    on    Pasture    Manured    with   Phosphates    and 

Potash,  Analyst  35,  246,  247. 

1905  Goldthwait,  J.  E.,  C.  F.  Painter,  R.  B.  Osgood,  and  F.  H.  McCrudden:  A 

Study  of  the  Metabolism  in  Osteomalacia,  Amer.  Jour.  Physiol.  14,  389- 
402. 

1910  Goldthwaite,  N.  E.:  Effects  of  the  Presence  of  Carbohydrates  upon  the 

Artificial  Digestion  of  Casein,  Jour.  Biol.  Chem.  7,  69-81. 

1905  Goliner:  Beitrag  zur  Wirkung  des  Lecithins,  Reichs-Medicinal-Anzeiger, 

Leipzig,  30,  7. 
1912     Goodman,  E.  H.:  Nuclease  in  Carcinoma,  Jour,  of  Exp.  Med.  15,  477-484. 

1906  Gordon,  Alfred:  The  Role  of  Organic  Phosphorus  in  the  Treatment  of 

Asthenic  Conditions  in  Various  Nervous  Diseases,  Medicine,  Detroit, 

12,  896-899. 
1898     Gossmann,  Heinrich:  Ueber  die  anorganischen  Bestandtheile  der  Bauch- 

speicheldriise  (Pankreas)  und  der  Niere,  Inaug.  Diss.,  Erlangen.  23  pp. 
1901     Gottstein,  Ernst:  Ueber  das  Verhalten  von  Kalcium  und  Magnesium  in 

einigen  Stoffwechselversuchen  mit  phosphorhaltigen  und  phosphorfreien 

Eiweisskorpern,  Inaug.  Diss.,  Breslau.       34  pp. 

1911  Goubau,  Fernand:  Sur  la  repartition  de  la  nucleohistone  dans  les  tissus 

des  mammiferes,  Bui.  Acad,  royale  de  med.  de  Belgique  25,  921,  946. 


PHOSPHORUS  METABOLISM  625 

1913  Goubau,  Fernand,  and  Maurice  Van  Goethem:  Etudes  sur  l'anaphylaxie 
par  les  nucleines.  I.  Memoire:  Action  de  l'injection  intraveineuse 
de  nucleine  et  de  nucleohistone  sur  la  circulation  chez  le  chien, 
Archives  internat.  de  physiol.  13,  289-304. 

1902-3  Gouin,  Andre,  and  Pierre  Andouard:  Nouvelles  recherches  sur  la  nutri- 
tion des  jeunes  bovides,  Bui.  Sta.  agron.  Loire-Inferieure,  66-100. 

1903  Gouin,  Andre,  and  Pierre  Andouard:  De  la  reaction  de  l'urine  des  bovides, 
Compt.  rend.  Soc.  de  biol.  55,  1600-1602. 

1905-6 Gouin,  Andre,  and  Pierre  Andouard:  Recherches  sur  l'alimentation  des 
jeunes  bovides,  Bui.  Sta.  agron.  Loire-Inferieure,  52-82. 

1907  Gouin,  Andre,  and  Pierre  Andouard:  Recherches  sur  l'alimentation  des 

jeunes  bovides,  Soc.  de  1'aliment.  rationelle  du  betail,  Compt.  rend. 
11th  congr.  11-28;  127-130. 

1908  Gouin,  Andre,  and  Pierre  Andouard:  Modes  d'elimination  des  phosphates 

dans  l'espece  bovine,  Compt.  rend.  Soc.  de  biol.  64,  133-135. 

1902  Gouraud,  F.  Xavier:  Courbe  d'elimination  des  phosphates  dans  la  pneu- 

monie  et  la  fievre  typhoide,  Ibid.  54,  373,  374. 

1903  Gouraud,  F.  Xavier:  Des  echanges  phosphores  dans  l'organisme  normal 

et  pathologique:  des  phosphaturies,  These  de  Paris.      135  pp. 
1894    Gourlay,  Frederick:  The  Proteids  of  the  Thyroid  and  the  Spleen,  Jour. 

of  Physiol.  16,  23-33. 
1912     Gouzien,  P.:  Le  beriberi  au  Tonkin,  Ann.  hyg.  med.  colon.  15,  445-491. 
1891    Graffenberger,  Louis:  Leber    die    Zusammensetzung    der    Kaninchen- 

knochen  im  hohen  Alter,  Landwirtsch.  Versuch.  Stat.  39,  115-126. 
1893    Graffenberger,  Louis:  1st  bei  der  Fiitterung  trachtiger  Tiere  die  Beigabe 

von  neutralem  phosphorsaurem  Calcium     zu     normal     beschaffenem 

Futter  fiir  die  Nachkommen  derselben  von  Nutzen?       Ibid.  41,  57-64. 
1905    Grandeau,  L.:  Les  maladies  des  os  et  ralimentation  phosphatee.  Rachit- 

isme.       Osteomalacia,  ou    cachexie     osseuse.        Jour,     d'agriculture 

pratique  120  (9  of  ser.  7),  726,  727. 
1902    Grandis,  V.,  and  Oscar  Copello:  Studi  sulla  composizione  chimica  delle 

ceneri  della  cartilagine  in  relazione    col    processo     di    ossificazione, 

Archivio  per  le  scienze  mediche  26,  175;  through     Jahresb.     u.     d. 

Fortschr.  d.  Thierchem.  32  (1902),  523,  524. 
1908    Granstrom,  E. :  Zur  Frage  iiber  den  Einfluss  der  Sauren  auf  den  Calcium- 

stoffwechsel  des  Pflanzenfressers,  Zeit.  physiol.  Chem.  58,  195-214. 
1911    Greenwald,  Isidor:  The  Effect  of  Parathyroidectomy  upon  Metabolism, 

Amer.  Jour.  Physiol.  28,  103-132. 
1913a  Greenwald,  Isidor :  Further  Metabolism  Experiments  upon  Parathyroidec- 

tomized  Dogs,  Jour.  Biol.  Chem.  14,  363-367. 
1913b  Greenwald,  Isidor:  On  the  Phosphorus  Content  of  the  Blood  of  Normal 

and  Parathyroidectomized  Dogs,  Ibid.  14,  369-379. 
1913c  Greenwald,  Isidor :  Bemerkungen  zu  der  Mitteilung  von  Raffaele  Paladino, 

"Untersuchungen   iiber   einige   Veranderungen   des   Stoffwechsels   bei 

Tieren  nach  Exstirpation  der  Schilddriise  und  der  Parathyroiden," 

Biochem.  Zeit.  54,  159,  160. 
1911    Gregersen,  J.  P.:  Untersuchungen  iiber  den  Phosphorstoffwechsel,  Zeit. 

physiol.  Chem.  71,  49-99. 
1913    Gregg,  D.:  Infantil  Beriberi  in  the  Philippines,  Boston  Med.  Surg.  J.  168, 

676-679;  title  through  Chem.  Abs.  7,  (1913),  2779. 


626  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1912  di  Gregorio,  Giovanni:     La  fitina     nel     ricambio     organico     fosforato 

fisiologico,  Bollett.   delle  sci.  med.  di  Bologna  83,  233-247;   through 

Zentralbl.  Biochem.  u.  Biophys.  14  (1912-13),  476. 
(1912)  Grijns,  G.:  Kritische  opmerkingen  over  "Jets  over  de  atiologie  van 

Polyneuritis  gallinarum,  in  verband  met  verzuurde  rijst."     (Etiology 

of  Polyneuritis  gallinarum  in  Connection  with  Sour  Rice)    Geneesk. 

Tijdschr.  Nederiandsch-Indie     52,  50;  through  Zentralbl.  Biochem.  u. 

Biophys.  14  (1912),  399. 
1903     Grimm:   Lecithin   und   seine   Bedeutung   fur   die   Kinderernahrung   als 

organische  Phosphorverbindung,  Vereinsblatt  der  pfalzischen  Aerzte 

19,  194-197. 
1905    Grindley,  H.  S.,  and  A.  D.  Emmett:  The  Chemistry  of  Flesh.  (Second 

paper.)       Improved  Methods  for  the  Analysis  of  Animal  Substances, 

Jour.  Amer.  Chem.  Soc.  27,  658-678. 

1913  Griniew,  D.  P.:  Les  lipoi'des  et  leur  teneur  en  phosphore  (dans  differents 

organes  et  tissus)  pendant  l'infection  tuberculeuse  chronique,  Archives 
des  sci.  biol.  17,  363-396. 
1913     Grosser,  Paul:  Ueber  den  Einflusz  des  Kochens  auf  das  physikalisch- 
chemische  Verhalten  von   Frauenmilch,  Kuhmilch,  und   Buttermilch, 
Biochem.  Zeit.  48,  427-431;  through  Chem.  Abs.  7  (1913),  1929. 

1912  Grosser,  Paul,  and  Joseph  Husler:  Ueber  das  Vorkommen  einer  Glycero- 
/        phosphatase  in  tierischen  Organen,  Ibid.  39,  1-5. 

1910     Grund,   Georg:   Organanalytische   Untersuchungen  liber   den   Stickstoff 

und  Phosphorstoffwechsel  und  ihre  gegenseitigen  Beziehungen,  Zeit. 

Biol.  54,  173-229. 
1912a  Grund,  Georg :  Ueber  die  chemische  Veranderung  des  Muskels  unter  dem 

Einfluss  der  Entartung,  Centralbl.  innere  Med.  33,  173-177;  through 

Chem.  Abs.  6  (1912),  1923. 
1912b  Grund,  Georg:  Zur  chemischen  Pathologie  des  Muskels,  Arch.  exp.  Path. 

u.  Pharm.  67,  393-407;  through  Chem.  Abs.  6  (1912),  2780. 

1913  Grund,  Georg:  Zur  chemischen  Pathologie  des  Muskels,  II.  Der  Einfluss 

der  Inaktivitatsatrophie   auf  die   Stickstoff-   und   Phosphorverteilung 
im  Muskel,  Ibid.  71,  129-138;  through  Chem.  Abs.  7  (1913). 

1892  Grundzach,  Ig.:  (Ueber  die  Asche  des  normal  en  Koths),  Gazeta  lekarska 

1892,  p.  48;  through  Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.,  22  (1892), 

311,  312. 
1903     Guerrini,  Guido:     Dell     azione     dei     nucleoproteidi     sulle     cellule     del 

parenchima  epatico,  La  Riforma  medica,  Palermo-Napoli  19,  707-709. 
1894     Gumlich:  Ueber  die  Aufnahme  der  Nucleine  in  den  tierischen  Organis- 

mus,  Zeit.  physiol.  Chem.  18,  508-512. 
1905     Gumpert,  E. :  Beitrag  zur  Kenntniss  des  Stickstoff-,  Phosphor-,  Kalk-,  und 

Magnesia-Umsatzes  beim  Menschen,  Med.  Klinik  1,  1037-1041. 
1883     Gurther,    Gustav:    Ueber    Veranderungen    im    Stoffwechsel    unter    dem 

Einfluss  der  Hypnose  und  bei  der  Paralysis  agitans,  Arch.  Psychiatrie 

14,  17-39. 

1893  Gusmitta,  Mario:  Sur  les  alterations  des  os  produites  par  l'inanition, 

Archives  ital.  de  biol.  19,  220-232. 
1862     Gusserow:     Beitrag  zur     Lehre     von    der     Osteomalacic,     Monatsschr. 

Geburtskunde  20,  19-25. 
1896-7  Gutinkov,  S. :  Zur  Lehre  von  der    chemischen     Zusammensetzung     des 

menschlichen  Gehirns,  Allgem.  Zeit.  Psychiat.  53,  270-329. 


PHOSPHORUS  METABOLISM  627 

1910  de  Haan,  J.:  On  the  Etiology  of  Beriberi,  Philippine  Jour,  of  Sci.,  B,  5, 
65-71. 

1907  Hamatainen,  Juho,  and  Waino  Helme:  Ein  Beitrag  zur    Kenntnis    des 

Eiweissstoffwechsels,  Skand.  Arch.  Physiol.  19,  182-200. 
1909    Haensel,  E.:  Ueber  den  Eisen-  und  Phosphorgehalt  unserer  Vegetabilien, 

Biochem.  Zeit.  16,  9-19. 
1912    Hagemann,  O.:  Beitrag  zur  Wirkung  der  Mineralsubstanzen  im  Tier- 

korper,  Arch.  ges.  Physiol.  146,  455-483. 
1900    Hahn,  M„,  and  L.  Geret:  Ueber  das  Hefe-Endotrypsin,  Zeit.  Biol.  40,  117- 

172. 

1895  Haiser,  F. :  Zur  Kenntniss  der  Inosinsaure,  Monatshef  te  Chem.  16, 190-206. 
1909    Haiser,  F.,  and  F.  Wenzel:  Ueber  Karnin  und  Inosinsaure,  Ibid.  30,  147- 

164;  377-386. 
1912    Hall,  L.  D.,  and  A.  D.  Emmett:    Relative    Economy,    Composition    and 
Nutritive  Value  of  the  Various  Cuts  of  Beef,  Univ.  111.  Agr.  Exp. 
Sta.  Bui.  No.  158.      233  pp. 

1896  Hall,  Winf.  S.:     Einige     Bemerkungen    iiber    die     Herstellung     eines 

kunstlichen  Futters,  Arch.  Anat.  u.  Physiol.,  142-153. 

1892  Halliburton,  W.  D.:  The  Proteids  of  Kidney  and  Liver  Cells,  Jour,  of 

Physiol.  13,  806-846. 

1893  Halliburton,  W.  D.:  The  Chemical  Physiology  of  the  Animal  Cell.  British 

Med.  Jour.  1,  501-506,  572-577,  627-632. 

1894  Halliburton,  W.  D.:  The  Proteids  of  Nervous  Tissue,  Jour.  Physiol.  15, 

90-107. 

1895  Halliburton,  W.  D.:  Nucleo-proteids.       Supplementary  paper,  Ibid.  18, 

306-318. 

1901a  Halliburton,  W.  D.:  The  Physiological  Effects  of  Extracts  of  Nervous 
Tissues,  Ibid.  26,  229-243. 

1901b  Halliburton,  W.  D.:  The  Chemical  Side  of  Nervous  Activity.  I.  The 
General  Composition  of  Nervous  Structures.  II.  Metabolism  in 
Nervous  Tissues.  III.  The  Chemical  Pathology  of  Hyperpyrexia,  and 
the  Chemical  Pathology  of  General  Paralysis  of  the  Insane.  IV.  The 
Chemistry  of  Nerve  Degeneration.  The  Croonian  Lectures  on  the 
Chemical  Side  of  Nervous  Activity.  1901,  London.  Through  Lancet, 
1901,  1,  1659,  1660;  1741,  1742. 

1905  Halliburton,  W.  D.:  Die  Biochemie  der  peripheren  Nerven,  Ergebnisse  d. 
Physiol.  4,  23-83. 

1894  Halliburton,  W.  D.,  and  T.  Gregor  Brodie:  Nucleo-albumins  and  Intra- 
vascular Coagulation,  Jour,  of  Physiol.  17,  135-173. 

1896  Halliburton,  W.  D.,  and  T.  Gregor  Brodie:  Action  of  Pancreatic  Juice  on 

Milk,  Ibid.  20,  97-106. 
1889     Halliburton,  W.  D.,  and  W.  M.  Friend:  The  Stromata  of  the  Red  Cor- 
puscles, Ibid.  10,  532-549. 

1908  Halpern,  Mieczyslaw:  Beitrag  zum  Hungerstoffwechsel,  Biochem.  Zeit. 

14,  134-142. 

1906-7  Hamill,  J.  Molyneux:  Observations  on  Human  Chyle,  Jour,  of  Physiol. 
35, 151-162. 

1877  Hammarsten,  Olof :  (Zur  Kenntnis  des  Caseins  und  der  Wirkung  des 
Labfermentes,)  Nova  Acta  Regiae  Soc.  Sci.  Upsaliensis  in  Memoriam 
Quattuor  Saec.  ab  Univ.  Upsaliensi  Peractorum.  1877.  Through 
Jahresb.  u.  d.  Fortschr.  d.  Thierchem.  7  (1877),  158. 


628  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1883    Hammarsten,  Olof :  Zur  Frage,  ob  das  Casein  ein  einheitlicher  Stoff  sei, 

Zeit.  physiol.  Chem.  7,  227-273. 
1885a  Hammarsten,  Olof:  Ueber  den  Gehalt  des  Caseins  an  Schwefel  und  iiber 

die  Bestimmung  des  Schwefels  in  Proteinsubstanzen,  Ibid.  9,  273-309. 
1885b  Hammarsten,  Olof:  Studien  iiber  Mucin  und  mucinahnliche  Substanzen, 

Arch.  ges.  Physiol.  36,  373-456. 
(1893)  Hammarsten,  Olof:  Till    kaunedomen     om     Nukleoproteiderun,     (Zur 

Kenntniss  der  Nucleoprote'ide),  Upsala  Lakareforenings  Forhandl.  Bd. 

28;  through  Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  23  (1893),  35-37. 
1894    Hammarsten,  Olof:   Zur  Kenntniss  der   Nucleoproteide,  Zeit.  physiol. 

Chem.  19,  19-37. 
(1895)  Hammarsten,  Olof:  Nagra  ord  om  olikheterna  mellan  qvinnomjolk  och 

komjolk,   Upsala    Lakareforenings    Forhandlinger,    Bd.    30;    through 

Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  25  (1895),  206,  207. 

1896  Hammarsten,  Olof:  Ueber  das  Verhalten  des  Paracaseins  zu  dem  Lab- 

enzyme,  Zeit.  physiol.  Chem.  22,  103-126. 

1901  Hammarsten,  Olof:  Untersuchungen  iiber  die  Gallen  einiger  Polarthiere. 

I.  Ueber  die  Galle  des  Eisbaren.      Erster  Abschnitt,  Ibid.  32,  435-466. 

1902  Hammarsten,  Olof:  Untersuchungen  iiber  die  Gallen  einiger  Polarthiere. 

I.  Ueber  die  Galle  des  Eisbaren.      II.  Abschnitt,  Ibid.  36,  525-555. 
1904     Hammarsten,  Olof:  Untersuchungen  iiber  die  Gallen  einiger  Polartiere. 

II.  Ueber  die  Galle  des  Moschusochsen,  Ibid.  43,  109-126. 

1905a  Hammarsten,  Olof:  Zur  Chemie  des  Fischeies,  Skand.  Arch.  Physiol.  17, 

113-132. 
1905b  Hammarsten,  Olof:  Zur  Chemie  der  Galle,  Ergebnisse  d.  Physiol.  4,  I. 

Abt.,  1-22. 
1908    Hammarsten,  Olof:  Zur  Frage  nach  der    Identitat    der    Pepsin-    und 

Chymosinwirkung,  Zeit.  physiol.  Chem.  56,  18-80. 
1911     Hammarsten,  Olof:  A  Text  Book  of  Physiological  Chemistry,  from  the 

Author's  7th  German  Edition. 
1881     Hammerbacher,  Friedr.:  Quantitative  Verhaltnisse  der  organischen  und 

unorganischen  Bestandtheile  des  menschlichen  gemischten  Speichels, 

Zeit.  physiol.  Chem.  5,  302-308. 
1863     Hammond,  William  A. :  On  the  Excretion  of  Phosphoric  Acid  through  the 

Kidneys,  Physiological  Memoirs,  Philadelphia,  1863,  29-41. 

1897  Hanai,  T.:  Physiological  Observations  on  Lecithin,  Tokyo  Imper.  Univ., 

Coll.  Agr.  Bui.,  2,  503-506. 
(1911)  Handovsky,  Hans,  and  Rich.  Wagner:  Ueber  einige  physikalisch-chem- 

ische  Eigenschaften  von     Lecithinemulsionen     und     Lecithineiweiss- 

mischungen,  Biochem.  Zeit.  31,  32-46;  through  Jahresb.  ii.  d.  Fortschr. 

d.  Thierchem.  41  (1911),  42. 
1912a  Hanes,  Frederick  M.:  Ueber  das  Vorkommen  und  die  Bedeutung  von 

anisotropen    Lipoiden    in    der  Leber  des    Huhnerembyros,    Centralbl. 

allgem.  Path.  23,  533-537;  through  Chem.  Abs.  6  (1912),  2627,  2628. 
1912b  Hanes,  Frederick  M.:  Lipoid  Metabolism  in  the  Developing  Chick  and  its 

Relation  to  Calcification,  Jour.  Exp.  Med.  16,  512-526;  through  Chem. 

Abs.  7  (1913),  1909. 
1906    Hannes,  Walther:  Resistenzerhohung  des  Peritoneums  gegen  Infektion 

mittels    Nukleinsaure,    eine   prophylaktische      Massnahme,       um    die 

Morbiditat  und  Mortalitat  nach  der  abdominalen  Radikaloperation  des 

gebarmutterkrebses  herabzusetzen,  Zentralbl.  Gynakol.  30,  681-692. 


PHOSPHORUS  METABOLISM  629 

1910  Harden,  Arthur,  and  Roland  V.  Norris:  The  Fermentation  of  Galactose 
by  Yeast  and  Yeast- juice  (Preliminary  Communication),  Proc.  Roy. 
Soc.  of  London  82  B,  645-649. 

1906  Harden,  Arthur,  and  William  John  Young:  The  Alcoholic  Ferment  of 

Yeast- juice,  I.  and  II.  Ibid.  77  B,  405-420;  78  B,  369-375. 
1908a  Harden,  Arthur,  and  William  John     Young:     The     Fermentation     of 
Mannose  and  Laevulose  by  Yeast- juice,  Proc.  of  the  Chem.  Soc.  24, 

115,  116. 
1908b  Harden,  Arthur  and  William  John  Young:  The  Alcoholic  Ferment  of 
Yeast-juice.      III.      Proc.  Roy.  Soc.  of  London  80  B,  299-311. 

1909  Harden,  Arthur,  and  William  John  Young:  The  Alcoholic  Ferment  of 

Yeast- juice.      Part  IV.      The  Fermentation  of  Glucose,  Mannose  and 

Fructose  by  Yeast- juice,  Ibid.  81  B,  336-347. 
1911b  Harden,  Arthur,  and  William  John  Young:  Ueber  die  Zusammensetzung 

der  durch  Hefepresssaft  gebildeten  Hexosephosphorsaure.       I.   and 

II.  Biochem.  Zeit.  32,  173-176;  177-188. 
1912    Harden,  Arthur,  and  William  John  Young:  Der  Mechanismus  der  alkohol- 

ischen  Garung,  Ibid.  40,  458-478;  through  Zeit.  Unters.  d.  Nahrungs-  u. 

Genussmittel  25  (1913),  456. 

1910  Harden,  J.:  (Neuere  Untersuchungen  liber  alkoholische  Garung),  Jour,  of 

the  Inst,  of  Brewing  1910,  623;  through  Zeit.  Unters.  d.  Nahrungs-  u. 
Genussmittel  23  (1912),  164,  165. 

1892  Hardin,  M.  B.:  I.  On  the  Available  Phosphoric  Acid  and  the  Water- 
Soluble  Potash  in  Cotton  Seed  Meal.  II.  On  the  Methods  of  Prepar- 
ing Solutions  of  Cotton  Seed  Meal  for  Precipitation  of  the  Phosphoric 
Acid.  III.  On  the  Occurrence  of  Meta-Phosphoric  Acid  and  Pyro- 
Phosphoric  Acid  in  Cotton  Seed  Meal,  So.  Carolina  Agr.  Exp.  Sta.  Bui. 
No.  8,  New  Series.      16  pp. 

1892  Harris,  V.  D.,  and  Wm.  J.  Gow:  Ferment  Actions  of  the  Pancreas  in 
Different  Animals,  Jour,  of  Physiol.  13,  469-492. 

1907  Hart,  E.  B. :  Variations  in  the  Amount  of  Casein  in  Cow's  Milk,  Twenty- 

fourth  Ann.  Rept.  Wis.  Agr.  Exp.  Sta.  111-116. 
1909     Hart,  E.  B. :  A  Volumetric  Method  for  the  Estimation  of  Casein  in  Cow's 

Milk,  Jour.  Biol.  Chem.  6,  445-451. 
1903     Hart,  E.  B.,  and  W.  H.  Andrews:  The  Status  of  Phosphorus  in  Certain 

Food  Materials  and  AnimaL  By-products,  with  Special  Reference  to  the 

Presence  of  Inorganic  Forms,  N.  Y.  Agr.  Exp.  Sta.  Bui.  238,  181-196; 

also  Amer.  Chem.  Jour.  30,  470-485. 
1909     Hart,  E.  B.,  E.  V.  McCollum,  and  J.  G.  Fuller:  The  Role  of  Inorganic 

Phosphorus  in  the  Nutrition  of  Animals,  Amer.  Jour.  Physiol.  23, 

246-277. 

1909  Hart,  E.  B.,  E.  V.  McCollum,  and  G.  C.  Humphrey:  The  Role  of  the  Ash 

Constituents  of  Wheat  Bran  in  the  Metabolism  of  Herbivora,  Ibid.  24, 
86-103;  also  Univ.  Wis.  Agr.  Exp.  Sta.,  Research  Bui.  No.  5. 

1910  Hart,  E.  B.,  and  W.  E.  Tottingham:  The  Nature  of  the  Acid-Soluble  Phos- 

phorus Compounds  of  Some  Important  Feeding  Materials,  Univ.  Wis. 
Agr.  Exp.  Sta.,  Research  Bui.  No.  9,  95-106;  also  Jour.  Biol.  Chem.  6, 
431-444. 
1901  Hartenberg,  P.:  La  lecithine  dans  la  therapeutique  des  affections  du 
systeme  nerveux,  11th  Congr.  des  med.  alienistes  et  neurologistes  de 
France,  p.  392. 


630  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1913  Hartwell,  Burt  L.:  The  Percentage  of  Total  Phosphorus  in  Flat  Turnips 
as  Influenced  by  the  Amount  Available  in  Soils,  Agr.  Exp.  Sta.  of  R.  I. 
State  Coll.,  Bui.  154,  121-148. 

1911  Hartwell,  Burt  L.,  and  Frederick  S.  Hammett:  The  Effect  of  Phosphorus 

Manuring  on  the  Amount  of  Inorganic  Phosphorus  in  Flat  Turnip 

Roots,  Jour.  Ind.  and  Engin.  Chem.  3,  831,  832. 
1910     Hartwell,  Burt  L.,  and  Wilhelm  B.  Quantz:  The  Phosphorus  of  the  Flat 

Turnip,  Proc.  Amer.  Soc.  Biol.  Chemists,  1909,  in  Jour.  Biol.  Chem.  7, 

xxxviii. 
1888     Hasebroek,  Karl:  Ueber  das  Schicksal  des  Lecithins  im  Korper,  und  eine 

Beziehung   desselben   zum   Sumpfgas   im   Darmcanal,   Zeit.   physiol. 

Chem.  12,  148-162. 
1903     Hatai,  Shinkishi:  The  Effect  of  Lecithin  on  the  Growth  of  the  White 

Rat,  Amer.  Jour.  Physiol.  10,  57-66. 
1854     Haubner:  Wissenschaftliche  und  praktische  Mittheilungen.  Ueber  einige 

dyskasische  Knochenkrankheiten,  Magazin  f.   d.   ges.   Thierheilk,  20, 

197-242. 
1867    Haubner:  Sitz.  Ber.  d.  Ges.  f.  Natur-  u.  Heilk.  zu  Dresden,  1867;  through 

Jahrbticher  der  in-  u.  auslandisch.  ges.  Med.  151  (1871),  138. 

1875  Haubner:  Rachitis  und  Tuberkulose  beim  Rinde  nach  Huttenrauchfutter, 

Jahresb.  d.  Ges.  f.  Nat.  u.  Heilk.  76,  115. 

1903  Hawk,  P.  B.:  On  the  Time  Relations  of  Proteid  Metabolism,  Amer.  Jour. 

Physiol.  10,  115-145. 
1905     Hawk,  P.  B.:  On  the  Influence  of  Copious  Water  Drinking,  Univ.  Penn. 
Med.  Bui.  March,  1905,  53  pp. 

1904  Hawk,  P.  B.,  and  Joseph  S.  Chamberlain:  A  Study  of  the  Variations  in 

the  Course  of  the  Nitrogen,  Sulphate  and  Phosphate  Excretion,  as 

Observed  in  Short  Periods  Following  a  Small  Increase  in  the  Proteid 

Ingested,  Amer.  Jour.  Physiol.  10,  269-289. 
1904     Hawk,  P.  B.,  and  William  J.  Gies:  The  Influence  of  External  Hemorrhage 

on  Chemical  Changes  in  the  Organism,  with  Particular  Reference  to 

Proteid  Catabolism,  Ibid.  11,  171-236. 
1860    Haxthausen,  Hugo  Augustus  Ab.:  Acidum  phosphoricum  urinae  et  excre- 

mentorum,  Inaug.  Diss.,  Halle. 
1886    Heckel,  Ed.,  and  Schlagdenhauffen:  Sur  la  presence  de  la  lecithine  dans 

les  vegetaux,  Compt.  rend.  Acad,  des  sci.  103,  388-390. 

1890  Heffter,  Arthur:  Lecithin  in  der  Leber,  Inaug.  Diss.,  Leipzig;  through 

Chem.  Centralbl.  1   (1891),  459. 

1891  Heffter,  Arthur:  Das  Lecithin  in  der  Leber  und  sein  Verhalten  bei  der 

Phosphorvergiftung,  Arch.  exp.  Path.  u.  Pharm.  28,  97-112. 

1903  Heffter,  Arthur:  Die  Ausscheidung  korperfremder  Substanzen  im  Harn. 

I.  Teil:  Anorganische  Verbindungen,  Ergebnisse  d.  Physiol.  2,  I.  Abt. 
95-129. 

1904  Heim,  Max.:  Klinische  Erfahrungen  mit  "Bioson,"  einer  Eiweiss-Eisen- 

Lecithin-Verbindung,  Berlin,  klin.  Wochenschr.   592-597. 

1912  Heiser,  V.  G.:  Beri-beri:  with  a  Suggestion  for  Governmental  Aid  in  its 

Eradication,  Med.  Record  81,  516,  517. 

1876  Heiss,  Ernst:  Kann  man  durch  Einfiihrung  von  Milchsaure  in  den  Darm 

eines    Thieres    den   Knochen   anorganischen    Bestandteile    entziehen? 
Zeit.  Biol.  12,  151-169. 


PHOSPHORUS  METABOLISM  631 

1873  Heitzmann,  C:  Ueber  kiinstliche  Hervorrufung  von  Rhachitis  und  Osteo- 
malacie,  Wien.  med.  Presse,  1873,  1035-1037;  Wien.  med.  Anzeiger  113 
(1873);  and  Wien.  med.  Jahrbiicher  1874. 

1894  Hempel,  Walther:  Die  Milchuntersuchungen  Professor  Dr.  Julius  Leh- 
mann's  Arch.  ges.  Physiol.  56,  558-578. 

1906  Henderson,  Lawrence  J.:  Equilibrium  in  Solutions  of  Phosphates,  Amer. 

Jour.  Physiol.  15,  257-271. 
1908a  Henderson,  Lawrence  J.:  A  Note  on  the  Union  of  the  Proteins  of  Serum 

with  Alkali,  Ibid.  21,  169-172. 
1908b  Henderson,  Lawrence  J.:  Concerning    the    Relationship    between    the 

Strength  of  Acids  and  their  Capacity  to  Preserve  Neutrality,  Ibid.  21, 

173-179. 
1908c  Henderson,  Lawrence  J.:  The  Theory  of  Neutrality  Regulation  in  the 

Animal  Organism,  Ibid.  21,  427-448. 
1909    Henderson,  Lawrence  J.:  On  the  Neutrality  Equilibrium  in  Blood  and 

Protoplasm,  Jour.  Biol.  Chem.  7,  29-35. 

1911  Henderson,  Lawrence  J. :  A  Critical  Study  of  the  Process  of  Acid  Excre- 

tion, Ibid.  9,  403-424. 
1913    Henderson,  Lawrence  J.:  The  Regulation  of  Neutrality  in  the  Animal 
Body,  Science,  N.  S.  37,  389-395. 

1907  Henderson,  L.  J.,  and  O.  F.  Black:  Concerning  the  Neutrality  of  Proto- 

plasm, Amer.  Jour.  Physiol.  18,  250-255. 

1908  Henderson,  L.  J.,  and  O.  F.  Black:  A  Study  of  the  Equilibrium  between 

Carbonic  Acid,  Sodium  Bicarbonate,  Mono-sodium  Phosphate,  and  Di- 

sodium  Phosphate  at  Body  Temperature,  Ibid.  21,  420-426. 
1903    Henderson,  Yandell,  and  Gaston  H.  Edwards:  Nuclein  Metabolism  in 

Lymphatic  Leukaemia,  Ibid.  9,  417-424. 
1897    Henriques,  Valdemar:  Ueber  die  reducirenden  Stoffe  des  Blutes,  Zeit. 

physiol.  Chem.  23,  244-257. 
1903    Henriques,  V.,  and  C.  Hansen:  Ueber  den  Uebergang  des  Nahrungsfettes 

in  das  Huhnerei  und  iiber  die  Fettsaure  des  Lecithins,  Skand.  Arch. 

Physiol.  14,  390-397. 

1889  Henry,  W.  A.:  Experiments  in  Pig  Feeding,  Sixth  Ann.  Rept.  Wis.  Agr. 

Exp.  Sta.,  pp.  6-41. 

1890  Henry,  W.  A. :  Feeding  Bone  Meal  and  Hard  Wood  Ashes  to  Hogs  Living 

on  Corn,  Univ.  Wis.  Agr.  Exp.  Sta.  Bui.  No.  25,  3-10. 
1719    Hensing,  Joh.  Th.:  Examen  chemicum  cerebri,  ex  eodemque  phosphorus 
singularis  omnia  inflammans,  Diss.,  Giessae;  through  Thudichum,  1901. 

1912  Herbst,  O.:  Beitrage  zur  Physiologie  des  Stoffwechsels  im  Knabenalter 

mit  besonderer  Berucksichtigung  einiger  Mineralstoffe,  Jahrb.  Kinder- 
heilk.  76,  Erganzungsh.,  40-130. 

1913  Herbst,  O.:  Calcium  und  Phosphor  beim  Wachstum  am  Ende  der  Kind- 

heit,  Zeit.  Kinderheilk.      7,  161-192. 
1900    Herlant,  Leon:  Untersuchungen  iiber  die  Nucleinsaure     aus     unreifer 

Lachsmilch,   aus   Kalbsthymus   und  aus   Hefe,  Arch.   exp.   Path.   u. 

Pharm.  44,  148-159. 
1902a  Herlitzka,  Amedeo,  and  Angeola  Borrino:  Ricerche  sull'  azione  chimico- 

fisiologica  dei  nucleohistoni  e  dei    nucleoproteidi,    Giornale    d.    reale 

Accad.  di  med.  di  Torino  65  (8  of  ser.  4),  351-357. 
1902b  Herlitzka,  Amedeo,  and  Angeola  Borrino:  Ricerche  sull'  azione  biochem- 

ica  di  alcuni  nucleohistoni  e  nucleoproteidi,  Lo  Sperimentale  56,  656- 

673. 


632  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1903    Herlitzka,  Amedeo,  and  Angeola  Borrino:  Recherches  sur  Taction  bio- 

chemique  de  quelques  nucleo-histones  et  nucleo-proteides,  Archives 

ital.  de  biol.  39,  1-17. 
1866    Hermann,  Ludimar:  Ueber  die  Wirkungsweise  einer  Gruppe  von  Giften. 

Anhang Ueber  das  Vorkommen  von  Protagon  im  Blute,  Arch.  Anat. 

u.  Physiol.  27-40;  also  Hoppe-Seyler's  Med.  Chem.  Unters.  1,  140. 
1912    Hermann,  Edmund,  and  Julius  Hermann:  Ueber  den  Lipoidgehalt  des 

Blutes  normaler  und  schwangerer  Frauen  sowie  neugeborener  Kinder, 

Biochem.  Zeit.  43,  47-55. 
1898    Herter,  C.  A.:  An  Experimental  Study  of  Fat  Starvation  with  Especial 

Reference  to  the  Production  of  Serous  Atrophy  of  Fat,  Jour,  of  Exp. 

Med.  3,  293-314. 
1897    Herxheimer:  Untersuchungen  iiber  die  therapeutische  Verwendung  des 

Kalkbrodes,  Berlin,  klin.  .Wochenschr.  34,  423-425. 
1897    Herzog,  Alois:  Flachs  und  Leinen,  Mitt.  d.  Verband.  d.  osterr.  Flachs- 

u.  Leinen-inter.  2,  Nr.  19;  through  Biedermann's  Centralbl.  f.  Agricul- 

turchem.  26  (1897),  570. 
1891     Hess,  E.,  and  Schaffer:  Ueber  den  Einfluss  des  verfiitterten  precipitierten 

phosphorsauren  Kalkes  auf    die    chemische    Zusammensetzung    der 

Milchasche,  Landwirtsch.  Jahrb.  d.  Schweiz  5,  76-78. 
1896    Hess,  N.,  and  E.  Schmoll:  Ueber  die  Beziehungen  der  Eiweiss  und  Para- 

nucleinsubstanzen   der   Nahrung   zur  Alluxurkorperausscheidung   im 

Harn,  Arch.  exp.  Path.  u.  Pharm.  37,  243-252;  also  Verhandl.  d.  14th 

Congr.  f.  innere  Med.  1896,  330-332. 

1909  Heubner,  W.:  Versuche  iiber  den     Phosphorumsatz     des     wachsenden 

Organismus,  Verhandl.  d.  26.  Versamml.  d.  Gesell.  Kinderheilk.,  81. 
Versamml.  d.  Gesell.  deut.  Naturforsch.  u.  Aerzte,  Salzburg  1909,  149- 
161. 

1910  Heubner,  W.:  Ueber    die    Phosphorausscheidung    eines    Neugeborenen, 

Arch.  exp.  Path.  u.  Pharm.  62,  253-257. 

1911  Heubner,  W.:  Versuche  iiber  den     Nahrungsphosphor,     Munch,     med. 

Wochenschr.  58,  II,  2543,  2544. 
1908    Heubner,  W.,  and  M.  Reeb:  Ueber  Menge  und  Verteilung  des  Phosphors 
in  einigen  Nahrungsmitteln,  Arch.  exp.  Path.  u.  Pharm.,  Sup.,  265- 
272. 

1905  Hewlett,  Albion  Walter:  The  Effect  of  the  Bile  upon  the  Ester-splitting 

Action  of  Pancreatic  Juice.      A  preliminary  communication.      Johns 

Hopkins  Hosp.  Bui.  16,  20,  21. 
1904a  Heymann,  Felix:  Zur  Einwirkung  der  Castration    auf    den    Phosphor- 

gehalt  des  weiblichen  Organismus,  Arch.  Gynakol.  73,  366-405. 
1904b  Heymann,  Felix:  Zur  Einwirkung  der  Kastration  auf  den  Phosphor- 

gehalt  des  weiblichen  Organismus,  Zeit.  physiol.  Chem.  41,  246-258. 
1896    Heymans,  J.  F.:  Recherches  experimental es  sur  l'inanition  chez  le  lapin, 

Arch,  de  pharmacodynamic  2,  315-353. 

1906  Hiestand,    O.:    Historische    Entwicklung   unserer   Kenntnisse    iiber    die 

Phosphatide.       Beitrage  zur  Kenntnis  der  pflanzlichen  Phosphatide, 
Diss.,  Zurich.      202  pp.;  through  Jahresb.  ii.  d.  Fortschr.  d.  Thierchem. 
37  (1907),  1141,  1142. 
1910     Highet,  H.  Campbell:  Beriberi  in  Siam,  Philippine  Jour,  of  Sci.,  B,  5, 
73-79. 


PHOSPHORUS  METABOLISM  633 

1885    Hiller,  E.:  Vergleichende  Untersuchungen  am     Skelett     eines     Vogels, 

Landwirtsch.  Versuch.  Stat.  31,  319-335. 
1887    Hills,  J.  L.:  Test  of  the  Availability  of  Bone  Meal  when  Fed  to  Milch 

Cows,  Vt.  Agr.  Exp.  Sta.,  Ann.  Rept.  1,  81-84. 
1894    Hills,  J.  L.:  The  Effect  of  Feeding  Bone  Meal  on  the  Character  of  the 

Milk,  Ibid.  7  (for  1893),  106,  107. 
1902    Hirschler,  August,  and  Paul  Terray:  Ueber  die  Bedeutung  der  anorgan- 

ischen  Salze  im  Stoffwechsel  des  menschlichen  und  tierischen  Organis- 

mus,  Math.  u.  naturwiss.  Ber.  aus  Ungarn  20  (Pub.  1905),  145-238; 

also  Math,  es  termeszettudom.      Ertesito,  1902,  20,  477-561,  through 

Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  32  (1902),  696,  697. 
1905    Hirschler,  August,  and  Paul  Terray:  Ueber  die  Bedeutung  der  anorgan- 

ischen  Salze  im  Stoffwechsel  des  Organismus,  Zeit.  klin.  Med.  57,  137- 

184. 
1902    His:  Zur  Phosphortherapie  bei  Osteomalacic,  Deut.  Arch.  klin.  Med.  73, 

546-554. 
1898-9  Hitzig,  Theod. :  Ueber  das  Verhalten  des  Harns  bei  Febris  intermittens, 

Ibid.  62,  358-389. 
1900     HIad'ik,  J.:     Zur  Kenntniss     der     Alkalescenzbestimmung     in     kleinen 

Blutmengen,  Zeit.  klin.  Med.  39,  194-201. 

1904  Hoennicke,  Ernst:  Zur  Theorie  Osteomalacic      Zugleich  zur  Lehre  von 

den  Krankheiten  der  Schilddriise.  Vorlaufige  Mittheilung,  Berlin, 
klin.  Wochenschr.  41,  1154-1156. 

1905  Hoennicke,  Ernst:  Ueber  das  Wesen  der  Osteomalacic      Ein  Beitrag  zur 

Lehre  von  den  Krankheiten  der  Schilddriise.  Nebst  Bemerkungen 
iiber  den  seelische  Zustand  bei  der  Knochenerweichung,  Halle.  78  pp. 
1910  Hosl,  J. :  Ueber  Unterschiede  in  der  tryptischen  und  peptischen  Spaltung 
des  Caseins,  Paracaseins  und  des  Paracaseinkalkes  aus  Kuh-  und 
Ziegenmilch,  Inaug.  Diss.,  Bern.      31  pp. 

1906  von  Hoesslin,  Heinrich:  Ueber  den  Abbau  des  Cholins  im  Tierkorper, 

Beitrage  z.  chem.  Physiol,  u.  Path.  8,  27. 

1909  von  Hoesslin,  Heinrich:  Experimentelle  Untersuchungen  zur  Physiologie 

und  Pathologie  des  Kochsalzwechsels.  Mit  Beitragen  iiber  die 
Wirkung  des  Kochsalzes  auf  dem  Umsatz  und  die  Ausscheidung  von 
Stickstoff  und  Phosphorsaure,  Zeit.  Biol.  53  (N.  F.  35),  25-92. 

1910  Ho ff strom,  K.  A.:  Eine  Stoffwechseluntersuchung  wahrend  der  Schwang- 

erschaft,  Skand.  Arch.  Physiol.  23,  326-420. 
1897    Hofmann,  K.  B.:  Augenblicker  Milchsauregehalt  des  Harnes  bei  Osteo- 

malakie,  Centralbl.  innere  Med.  18,  329-333. 
1873     Hofmeister,  V.:  Ein  Futterungsversuch  mit     Lammern,     Landwirtsch. 

Versuch.  Stat.  16,  126-183. 

1910  Holsti,  Oesten:  Zur  Kenntniss  des  Phosphorumsatzes  beim  Menschen, 

Skand.  Arch.  Physiol.  23,  143-153. 
1890     D'Hont,  Fred.:  Contribution  a  l'etude  du  lait,  Courtrai.      6  pp. 

1911  Hoobler,  B.  Raymond:  The  Role  of  Mineral  Salts  in  the  Metabolism  of 

Infants,  Amer.  Jour,  of  Diseases  of  Children  2,  107-140. 

1912  Hoobler,  B.  Raymond:  Mineral  Salts  and  Their  Relation  to  the  Dietary  of 

Infants  and  Young  Children,  Archives  of  Pediat.  29,  208-214. 

1911     Hooper,  D.:  Phosphorus  in  Indian  Foodstuffs,  J.  Proc.  Asiatic  Soc.  of 

Bengal  7,  313-322;  through  Chem.  Abs.  6  (1912),  1638,  1639. 


634  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1897    Hoover,  G.  F.,  and  Torald  Sollmann:  A  Study  of  Metabolism  During 

Fasting  in  Hypnotic  Sleep,  Jour.  Exp.  Med.  2,  405-411. 
1912    Hopkins,  F.  G.:  Feeding  Experiments  Illustrating  the  Importance  of 

Accessory  Factors  in  Normal  Dietaries,  Jour,  of  Physiol.  44,  425-460. 
1912    Hopkins,  F.  G.:  and  Allen  Neville:  A  Note  Concerning  the  Influence  of 

Diets  upon  Growth,  Biochem.  Jour.  7,  97-99. 
1862    Hoppe,  Ernst  Felix  Immanuel   (Hoppe-Seyler) :  Untersuchungen  iiber 

die  Constitution  des  Zahnschmelzes,  Arch.  path.  Anat.  u.  Physiol.  24, 

13-32. 
1866a  Hoppe,  Ernst  Felix  Immanuel  (Hoppe-Seyler):  Beitrage  zur  Kenntniss 

der  Constitution  des  Blutes,  Med.  chem.  Untersuch.,  133-150. 
1866b  Hoppe,  Ernst  Felix  Immanuel  (Hoppe-Seyler):  Ueber  einige  Bestand- 

theile  der  Maiskorner,  Ibid.,  162,  163. 
1878-9  Hoppe,  Ernst  Felix  Immanuel    (Hoppe-Seyler) :    Ueber    Lecithin    und 

Nuclein  in  der  Bierhefe,  Zeit.  physiol.  Chem.  2,  427-429. 
1907a  Hoppe,  Joseph:  Die  Verwendung    des    Nukleogens    bei    nervenkranken 

Kindern,  Therapie  der  Gegenwart  48  (N.  F.  9),  501-503. 
1907b  Hoppe,  Joseph:  Ein  Beitrag  zur  Kenntnis  des  Mineral- Stoffwechsels  der 

Idioten,  Zeit.  Erforsch.  u.  Behandl.  d.  jugendlichen  Schwachsinns  1, 

29-35. 
1889     Horbaczewski,  J.:  Untersuchung  iiber  die  Entstehung  der  Harnsaure  im 

Saugethierorganismus,    Sitzungsber.    d.    Math,    -naturw.      Classe     d. 

Kaiserlich.  Akad.  d.  Wissensch.  (Vienna),  98,  Abt.  3,  301-318. 
1891a  Horbaczewski,  J.:  Beitrage  zur  Kenntniss  der  Bildung  der  Harnsaure 

und  der  Xanthinbasen,   sowie   der  Entstehung   der  Leukocytosen  im 

Saugethierorganismus,  Ibid.  100,  Abt.  3,  78-132. 
1891b  Horbaczewski,  J.:  Beitrage  zur  Kenntniss  der  Bildung  der  Harnsaure 

und  der  Xanthinbasen,  sowie  der  Entstehung     der     Leukocytose     im 

Saugethierorganismus,  Monatshefte  Chem.  12,  221. 

1893  Horbaczewski,  J.:  Bemerkungen  zum  Vortrage  des  Hrn.  Alb.  Kossel.: 

"Ueber  Nucleisaure."      Arch.  Anat.  u.  Physiol.,  physiol.  Abt.,  109-115. 
1907     Horner,  Oskar:  Zum  Verhalten  des  Phytins  im  Organismus,  Biochem. 

Zeit.  2,  428-434. 
1906     Hotz,  Gerhard:  Phosphorsaure-  und  Kalkstoffwechsel  bei  Osteomalacie 

unter  dem  Einfluss  der  Phosphortherapie,  Diss.,  Basel.  32  pp.;  also 

Zeit.  exp.  Path.  u.  Ther.  3,  605-632. 

1894  Houdet,  V.:  Contribution  a  Tetude  du  colostrum  de  la  vache,  Annales  de 

lTnst.  Pasteur  8,  506-513. 
1901     Huchard,  H. :  L'emploi  therapeutique  de  la  lecithine,  Jour,  des  praticiens, 

439-441. 
1899a  Hugounenq,  L.:  Recherches  sur  la  statique  des  elements  mineraux  et 

particulierement  du  fer  chez  le  foetus     humain,     Jour,     de     physiol. 

et  de  path,  gener.  1,  703-711;  also  Compt.  rend.  Soc.  de  biol.  51,  337, 

338;  and  Compt.  rend.  Acad,  des  sci.  128,  1054-1.056. 
1899b  Hugounenq,  L.:  La  composition  minerale  de  l'enfant  nouveau-ne  et  la 

loi  de  Bunge,  Compt.  rend.  Acad,  des  sci.  128,  1419-1421;  also  Compt. 

rend.  Soc.  de  biol.  51,  523-525. 
1900     Hugounenq,  L.:  La  composition  minerale  de     l'organisme     de     l'enfant 

nouveau-ne,  Jour,  de  physiol.  et  de  path,  gener.  2,  1-5. 
1905a  Hugounenq,  L.,  and  A.  Morel:  Recherches  sur  l'hematogene,  Compt.  rend. 

Acad,  des  sci.  140,  1065-1067. 


PHOSPHORUS  METABOLISM  635 

1905b  Hugounenq,  L.,  and  A.  Morel:  Recherches  sur  la  formation  de  l'hemo- 
globin  chez  l'embryon,  Ibid.  141,  848,  849. 

1901a  Huiskamp,  W.:  Ueber  die  Eiweisskorper  der  Thymusdriise,  Zeit.  physiol. 
Chem.  32,  145-196. 

1901b  Huiskamp,  W.:  Ueber  die  Elektrolyse  der  Salze  des  Nucleohistons  und 
Histons,  Ibid.  34,  32-54. 

1903  Huiskamp,  W.:  Beitrage  sur  Kenntniss  des  Thymusnucleohistons,  Ibid. 
39,  55-72. 

(1912)  Hulshoff-Pol,  D.  J.:  Jets  over  de  atiologie  van  Polyneuritis  gallinarum 
in  verband  met  verzuurde-rijstvoeding.  (Etiology  of  Polyneuritis  gall- 
inarum in  Hens  in  Connection  with  the  Feeding  of  Sour  Rice),  Geneesk. 
Tijdschr.  Nederlandsch-Indie  52,  11;  through  Zentralbl.  Biochem.  u. 
Biophys.  14  (1912),  399. 

1867  Huppert,  H.:  Analyse  eines  osteomalacischen  Knochens,  Archiv  der  Heil- 
kunde  8,  345-351. 

1901-2  Hutchison,  Robert,  and  J.  J.  R.  MacLeod:  A  Contribution  to  our  Know- 
ledge of  the  Chemistry  of  Red  Bone  Marrow,  Jour,  of  Anat.  and 
Physiol.  36,  292-295. 

(1911)  Ibrahim,  J.:  Kaseinklumpen  im  Kinderstuhl  im  Zusammenhang  mit 
Rohmilchernahrung,  Monatsschr.  f.  Kinderheilk.  10,  55-64;  through 
Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  41  (1911),  298. 
1901  Iljin,  M.  D.:  (Der  Einfluss  der  organischen  Phosphorverbindungen  auf 
die  Stickstoffablagerung  im  menschlichen  Korper),  Russkij  Wratsch 
5,  574;  through  Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  31  (1901),  733. 

1906  Iljin,  M.  D.:  (Gegenseitige  Beziehungen  der  Lecithine,  des  Phytins  und 

der  Nucleinsauren  auf  der  Grundlage  ihrer  Konstitution.)      Russkij 
Wratsch  1906,  5,  390-392;  through  Biochem.  Centralbl.  5  (1906-7),  534- 
538. 
1897    Imbert,  M.,  and  G.  Belugon:     Chaleur     de    neutralisation     de     l'acide 
glycerophosphorique,  Compt.  rend.  Acad,  des  sci.  125,  1040-1042. 

1907  Ingle,  Herbert:  Osteoporosis  in  Animals,  Jour,  of  Compar.  Path,  and 

Ther.  20,  35-48. 

1908  Ingle,  Herbert:  The  Mineral  Constituents  of  Foods,  Jour,  of  Agr.  Sci. 

3,  I,  22-31. 

1909  Ingle,  Herbert:  The  Importance  of  the  Mineral  Constituents  of  Foods, 

Jour,  of  the  Roy.  Inst,  of  Public  Health  17,  736-747." 

1904  Inouye,  Katsuji:  Ueber  das  Vorkommen  einer  Lavulinsaure  bildenden 

Atomgruppe  in  Nucleinsauren,  Zeit.  physiol.  Chem.  42,  117-120. 

1906  Inouye,  Katsuji:  Ueber  die  Nucleinsaure  aus  den     Spermatozoen     des 

Hamo,  Ibid.  48,  181-184. 

1905  Inouye,  Katsuji,  and  Y.  Kotake:  Ueber  die  Darmnucleinsaure,  Ibid.  46, 

201-205. 
(1911)  Isaac:  Das  Jodocitin,  Med.     Klinik     (Berlin)     7, .  1541-1545;     through 

Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  41.(1911),  890. 
1903     Iwanoff,  Leonid:  Ueber  die  fermentative  Zersetzung  der  Thymonuclein- 

saure  durch  Schimmelpilze,  Ibid.  39,  31-43. 

1907  Iwanoff,  Leonid:  Ueber  die  Synthese  der  Phosphoorganischen  Verbind- 

ungen  in  abgetoteten  Hefezellen,  Ibid.  50,  281-288. 
1900    Jackson,  Holmes  C:  On  the  Phosphorus  Content  of  the  Paranuclein  from 
Casein,  Amer.  Jour.  Physiol.  4,  170-177. 


636  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1906    Jacob,  Ludwig:     Futterungsversuche    mit    einer    aus    den    einfachen 
Nahrungsstoffen  zusammengesetzten  Nahrung  an  Tauben  und  Ratten, 

Zeit.  Biol.  48  (N.  F.  30),  19-62. 
1898    Jacob,  Paul,  and  Peter  Bergell:  Ueber    den     Einfluss     nucleinhaltiger 

Nahrung  auf  Blut  und  Stoffwechsel  unter  besonderer  Beriicksichtigung 

des  Phosphorsaurestoffwechsels,  Zeit.  klin.  Med.  35,  171-211. 
1902    Jaeckel,  Hermann:  Ueber  die  Zusammensetzung  des  menschlichen  Fettes. 

Ein  Beitrag  zur  Analyse  der  Fette,  Zeit.  physiol.  Chem.  36,  53-84. 
1902    Jagerroos,  B.  H.:  Studien  iiber  den  Eiweiss-,  Phosphor-  und  Salzumsatz 

wahrend  der  Graviditat,  Arch.  Gynakol.  67,  517-590. 
1897    de  Jager,  L.:  Over  de  werking  van  labferment,  Nederl.  Tijdschrift  voor 

Geneeskunde   (2r.)   33,  253-267;  through  Jahresb.  ii.  d.  Fortschr.  d. 

Thierchem.  27  (1897),  276-279. 
1910    de  Jager,  L.:  Ueber  den  Gehalt  des  Harnes  an  Ammoniak,  Phosphor- 

saure,  sauren  Korpern  und  Aminosauren,  Zentralbl.  ges.  Physiol,  u. 

Path,  des  Stoffwechsels  11  (n.  s.  5),  241-255. 

1876  von  Jaksch,  Rudolf:  Ueber  das  Vorkommen  von  Nuclein  im  Menschen- 

gehirn,  Arch.  ges.  Physiol.  13,  469-473. 
1885     von  Jaksch,  Rudolf:  Ueber  Acetonurie  und  Diaceturie.      156  pp. 
1910    Janin:  Sur  une  theorie  phosphatique  du  beriberi,  Soc.  de  med.  et  d'hyg. 

tropicale,  Caducee  1910,  No.  2,  p.  25;  through  Arch.  Schiffs-  u.  Tropen- 

hygiene  15  (1911),  36. 

1877  Jarisch,  Adolf:  Untersuchungen  iiber  die  Bestandtheile  der  Asche  des 

Blutes,  Wien.  med.  Jahrbucher,  39-64. 
1910    Jebbink,  G.  J.:  Over  het  nucleingehalte  van  menschelijk  voedsei  en  voor- 

al  van  Indische  versnaperingen,  Diss.  Amsterdam.      124  pp.  Through 

Jahresb.  u.  d.  Fortschr.  d.  Thierchem.  40,  640. 
1912    Jegorow,  M.  A.:  Zur  Kenntniss  der  Eigenschaften  des  Phytins,  Biochem. 

Zeit.  42,  432-439. 
1899    Jemma,  R.:  Sulla  digestione  artificiale  del  latte,  Clinica  med.  italiana 

1899,  No.  6;  through  Centralbl.  innere  Med.  21  (1900),  671,  672. 
1888    Jenkins,  E.  H.:  Observations  on  the  Chemical  Composition  of  Certain 

Grasses,  Conn.  Agr.  Exp.  Sta.  Rept.  for  1888,  Pt.  II,  100-102. 
1904    Jensen,  Orla:  Ueber  den  Einfluss  der  Mineralbestandteile  des  Futters  auf 

Milch,  Molkerei-Zeitung  14,  505-508;  517-519. 
1905a  Jensen,  Orla:  De  l'influence  des  elements  mineraux  du  fourrage  sur  le 

lait,  Revue  gener.  du  lait  4,  273-285;  297-306. 
1905b  Jensen,  Orla:  Lasst  die  Milch  sich  in  ihrer  Zusammensetzung  durch  das 

Futter  beeinflusst?     Landwirtsch.  Jahrb.  d.  Schweiz,  1905,  Separate 

abdruck;  through  Biedermann's  Centralbl.     f.    Agricultur-Chem.     35 

(1906),  285-287. 
1905-6  Jensen,  Orla:  La  composition  du  lait    peut-elle     etre    influencee    par 

l'aff ourragement  ?     Revue  gener.  du  lait  5,  103-110;  121-128;  152-161; 

178-185;  198-205. 
1897-8  Jerome,  William  J.  Smith :  The  Formation  of  Uric  Acid  in  Man,  and  the 

Influence  of  Diet  on  its  Daily  Output,  Jour,  of  Physiol.  22,  146-158. 
1899    'Jerome,  William  J.  Smith:  Further  Proofs  of  the  Origin  of  Uric  Acid 

from  Nuclein-compounds  and  Derivatives,  Ibid.  25,  98-103. 
1887a  Jolly  L.:  Sur  les  phosphates  et  leurs  fonctions  dans  les  etres  vivants, 

Paris.       Archives  gener.  de  med.  1,  627. 


PHOSPHORUS  METABOLISM  637 

1887b  Jolly,  L. :  Les  phosphates.      Leurs  f onctions  chez  les  etres  vivants,  Paris. 

578  pp. 
1898     Jolly,  L.:  Recherches  sur  le  phosphore  organique,  Compt.  rend.  Acad,  des 

sci.  126,  531-533. 
1904a  Jones,  Walter. :  Ueber  das  Enzym  der  Thymusdruse,  Zeit.  physiol.  Chem. 

41,  101-108. 
1904b  Jones,  Walter:  On  the  Enzyme  of  the  Suprarenal  Gland,  Proc.  Amer. 

Physiol.  Soc,  in  Amer.  Jour.  Physiol.  10,  XXV. 
1904c  Jones,  Walter:  Ueber  die  Selbstverdauung  von  Nucleoproteiden,  Zeit. 

physiol.  Chem.  42,  35-54. 

1905  Jones,  Walter:  Ueber  das  Vorkommen  der  Guanase  in  der  Rindermilz 

und  ihr  Fehlen  in  der  Milz  des  Schweines,  Ibid.  45,  84-91. 
1908.  Jones,  Walter:  On  the  Identity  of  the  Nucleic  Acids  of  the  Thymus, 

Spleen  and  Pancreas,  Jour.  Biol.  Chem.  5,  1-26. 
1910     Jones,  Walter:  Ueber  die  Beziehung  der  aus  wasserigen  Organextrakten 

gewonnenen  Nucleinfermente  zu  den  physiologischen  Vorgangen  im 

lebenden  Organismus,  Zeit.  physiol.  Chem.  65,  383-388. 
1911a  Jones,  Walter:  Concerning  Nucleases,  Jour.  Biol.  Chem.  9,  129-137. 
1911b  Jones,  Walter:  On  the  Physiological  Agents  Which  are  Concerned  in  the 

Nuclein  Fermentation,  with  Special  Reference  to  Four  Independent 

Desamidases,  Ibid.  9,  169-180. 
1912    Jones,  Walter:  On  the  Formation  of  Guanylic  Acid  from  Yeast  Nucleic 

Acid,  Ibid.  12,  31-35. 

1906  Jones,  Walter,  and  C.  R.  Austrian:  Ueber  die  Verteilung  der  Fermente 

des  Nucleinstoffwechsels,  Zeit.  physiol.  Chem.  48,  110-129. 
1907a  Jones,  Walter,  and  C.  R.  Austrian:  On  Thymus  Nucleic  Acid,  Jour.  Biol. 

Chem.  3,  1-10. 
1907b  Jones,  Walter,  and  C.  R.  Austrian:  On  the  Nuclein  Ferments  of  Embryos, 

Ibid.  3,  227-232. 

1904  Jones,  Walter,  and  C.  L.  Partridge:  Ueber  die  Guanase,  Zeit.  physiol. 

Chem.  42,  343-348. 
1914    Jones,  Walter,  and  A.  E.  Richards:  The  Partial  Enzymatic  Hydrolysis  of 

Yeast  Nucleic  Acid,  Jour  Biol.  Chem.  17,  71-80. 
1908    Jones,  Walter,  and  L.  G.  Rowntree:  On  the  Guanylic  Acid  of  the  Spleen, 

Ibid.  4,  289-295. 
1902    Jones,  Walter,  and  G.  H.  Whipple:  The  Nucleoproteid  of  the  Suprarenal 

Gland,  Amer.  Jour.  Physiol.  7,  423-434. 

1905  Jones,  Walter,  and  M.  C.  Winternitz:  Ueber  die  Adenase,  Zeit.  physiol. 

Chem.  44,  1-10. 
1885-6  Jordan,  W.  H.:  The  Manure  Residue  of  Corn  Meal  and  of  Cotton-Seed 
Meal,  Ann.  Rept.  Maine  Agr.  Exp.  Sta.  for  1885-6,  42-46. 

1906  Jordan,  W.  H.,  E.  B.  Hart,  and  A.  J.  Patten:  A  Study  of  the  Metabolism 

and   Physiological   Effects   of   Certain   Phosphorus   Compounds   with 

Milch  Cows,  Amer.  Jour.  Physiol.  16,  268-313;  also  N.  Y.  Agr.  Exp. 

Sta.,  Tech.  Bui.  1,  59  pp. 
1904    Joulie,  H.:  L'acide  phosphorique  dans  Palimentation  des  herbivores,  Rev. 

agr.  Reunion  10,  84-86. 
1912    Juchler,  Theodor:  Die  mineralischen  bestandteile  der  Bauchspeicheldriise, 

Inaug.  Diss.,  Zurich.    24  pp.    Through  Zeit.  Kinderheilk.  Referate  4 

(1913)  166,  167. 


638  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1868    Jiidell,  Gustav:  Zur  Blutanalyse,  Hoppe-Seyler's  Med.   Chem.  Unters. 

386-393. 
1913     Jundell,  I. :  Untersuchungen  iiber  den  Stoffwechsel  bei  der  Dyspepsie  und 

der  alimentaren  Intoxikation,  Zeit.  Kinderheilk.     Originalien  8,  235- 

290. 
1911     Juschtschenko,   A.:   Die    Schilddriise   und   die   fermentativen   Prozesse, 

Zeit.  physiol.  Chem.  75,  141-168. 
1911     Juschtschenko,  A.  J.:  "Ueber  den  Nucleasegehalt  verschiedener  Organe 

des  Menschen  und  der  Tiere,  Biochem.  Zeit.  31,  377-384. 
1913     Juschtschenko,  A.  S.:  Zur  Physiologie  der  Schilddriise:  Gehalt  an  Phos- 
phor, Stickstoff  und  Lipoiden  bei  Thyroidektomierten  Tieren,  Ibid.  48, 

64-85. 
1906     Kaas,  Karl:  Ueber  den  Phosphorgehalt  von  Huhnereiweiss,  Monatshefte 

Chem.  27,  403-409. 
1910     Kajiura,  S.,  and  O.  Rosenheim:  A  Contribution  to  the  Etiology  of  Beri- 
beri, Jour,  of  Hyg.  10,  49-55. 
1907a  Kalaboukoff,  and  Emile  F.  Terroine:  Sur  l'activation  des  ferments  par 

la  lecithine.       I.  Action  de  la  lecithine  sur  la  lipase  pancreatique, 

Compt.  rend  Soc.  de  biol.  63,  372-374. 
1907b  Kalaboukoff,  and  Emile  F.  Terroine:  Sur  l'activation  des  ferments  par 

la  lecithine.       II.  Action  de  la  lecithine  sur  les  lipases  gastrique  et 

intestinale,  Ibid.  63,  617-619. 
1907c  Kalaboukoff,  and  Emile  F.  Terroine:  Sur  l'action  de  la  lecithine  sur  les 

ferments.       III.  Action  de  l'ovolecithine  sur  l'amylase,  la  trypsine  et 

le  lab,  Ibid.  63,  664-666. 
1909    Kalaboukoff,  and  Emile  F.  Terroine:  Action  du  sue  pancreatique  et  des 

sels  biliares  sur  l'ovolecithine,  Ibid  66,  176-178. 
1897     Kalinin,  A.  A.:  Untersuchungen  iiber  die  Ausscheidung  von  Kohlensaure, 

Stickstoff  und  Phosphor  und  den  Sauerstoffverbrauch  in  der  Latenz- 

periode  des  Fiebers  bei  Kaninchen  und  Hunden  nach  subcutaner  Infec- 
tion  mit   Bouillonculturen   von   Pyocyaneus-   und   Diphtheriebacillen, 

Centralbl.  allgem.  Path.  u.  path.  Anat.  8,  518-524. 
1913     Kaminer,  Gisa,  and  Ernst  Mayerhofer:  Ueber  den  klinischen  Wert  der 

Bestimmung     des     anorganischen   Phosphors  im   Harne  unnaturlich 

ernahrter  Sauglinge,  Zeit.  Kinderheilk.    8,  24-49. 
1867    Karmrodt,  C:  Untersuchungen  von  Wiesenheu,  Zeit.  des  landwirtsch. 

Vereins  f.  Rheinpreussen  35,  376-378;  through  Jahresb.  f.  Agricultur- 

chem.  11  &  12  (1868-9),  488  and  545. 
1881     Kassowitz,  Max:  Die  normale  Ossification  und  die  Erkrankungen  des, 

Knochensystems  bei  Rachitis  und     hereditarer     Syphilis.       I.  Theil: 

Normale  Ossification,  Wien.  med.  Jahrbucher,  1881. 
1884a  Kassowitz,  Max:  Die  normale  Ossification  und  die  Erkrankungen  des 

Knochensystems  bei  Rachitis  und  hereditarer   Syphilis.       II.   Theil: 

Rachitis.       1.  Abtheilung.       2.  Abtheilung.       Die  Pathogenese  der 

Rachitis,   Ibid.   1884. 
1884b  Kassowitz,  Max:  Die  Phosphorbehandlung  der  Rhachitis,  Zeit.  klin.  Med. 

7,  36-74;  93-139. 
1886     Kassowitz,  Max:  Die  Symptome  der  Rachitis  auf  anatomischer  Grund- 

lage  bearbeitet.      I.  Abtheilung.  Verbildungen  und  Funktionsstorung- 

en  der  Extremitaten,  Leipzig.       130  pp. 
1890     Kassowitz,  Max:  Zur  Theorie  und  Behandlung  der  Rachitis,  Beitrage  z. 

Kinderheilk.       1890,  book  1,  106-149. 


PHOSPHORUS  METABOLISM  639 

1901  Kassowitz,  Max:  Zur  Theorie  der  Rachitis,  Wien.  med.  Wochenschr.  51, 

1753-1758';  1807-1813;  1857-1860. 

1910  Kassowitz,  Max:     15.    Vorlesung:  Knochenwachstum  und  Rachitis  (15 

pp.);  16.  Vorlesung:  Die  Ursachen  der  Rachitis  (14  pp.);  17.  Vor- 
lesung: Vorbeugung  und  Behandlung  der  Rachitis  (18  pp.),  Praktische 
Kinderheilkunde,  in  36  Vorlesungen,  Berlin. 

1912  Kassowitz,  Max:  Ueber  Rachitis.      II.  Osteochondritis  rachitica,  Jahrb. 

Kinderheilk.      75  (25  of  ser.  3),  194-212;  334-349;  489-505;  581-600. 

1913  Kassowitz,  Max:  Weitere  Beitrage  zur  Rachitisfrage.       III.  Phosphor 

und  Lebertran,  Deut.  med.  Wochenschr.  39  II,  1716-1720. 

1909  Kastle,  Joseph  H.,  and  Norman  Roberts:  The  Chemistry  of  Milk,  in 
"Milk  and  Its  Relations  to  the  Public  Health,"  Bui.  No.  56,  Hygienic 
Lab.  of  the  Pub.  Health  and  Marine-Hosp.  Service  of  U.  S.,  313-425. 
373  refs. 

1908  Katayama,  T.:  Ueber  die  Zusammensetzung  der  Kuhmilch  verschieden- 
er  Rassen  mit  besonderer  Berucksichtigung  ihres  Kalk-  und  Phos- 
phorsauregehaltes,  Landwirtsch.  Versuch.  Stat.  69,  342-358. 

1896  Katz,  Julius:  Die     mineralischen     Bestandtheile     des     Muskelfleisches, 

Archiv  ges.  Physiol.  63,  1-85. 

1902  Kauf  mann,-  Martin :  Ein  Beitrag  zur  Frage  der  Fleischmast,  Centralbl. 

Stoffwechsel-  und  Verdauungs-krankheit,  3,  239-241. 

1902  Kauf  mann,  M.,  and  L.  Mohr:  Beitrage  zur  Alloxurkorperfrage  und  zur 

Pathologie  der  Gicht,  Deut.  Arch.  klin.  Med.  74,  141-162;  348-369;  586- 
614. 

1903  Kauf  mann,  M.,  and  L.  Mohr:  Ueber  Eiweissmast,  Berlin,  klin.  Wochen- 

schr. 8,  161-163. 
1902     Kaup,  Igo :  Ein  Beitrag  zu  der  Lehre  vom  Einflusse  der  Muskelarbeit  auf 

den  Stoffwechsel,  Zeit.  Biol.  43,  221-255. 
1856a  Kaup,  Wilhelm:  Beitrage  zur  Physiologie  des  Harnes.       2.  Ueber  die 

Auf saugung  von  Harnbestandtheilen  in  der  Blase,  Arch,  physiol.  Heilk. 

125-164. 
1856b  Kaup,  Wilhelm:  Beitrage  zur  Physiologie  des  Harnes,  Ibid.  556-566. 

1911  Kaupp,  B.  F.:  The  Effect  of  Nuclein  on  the  Blood,  Amer.  Veterinary 

Review,  40,  306-315. 

1897  Keller,  Arthur:  Zur  Kenntniss   der   Gastroenteritis  im   Sauglingsalter, 

Ammoniak-Ausscheidung,  Jahrb.  Kinderheilk.   44,  25-52. 

1898  Keller,  Arthur:  Phosphorstoffwechsel  im  Sauglingsalter,  Zeit.  klin.  Med. 

36,  49-76. 
1900a  Keller,  Arthur:  Organische  Phosphorverbindungen  in  Sauglingsharn,  ihr 

Ursprung  und  ihre  Bedeutung  fur  den  Stoffwechsel.     Zeit.     physiol. 

Chem.  29,  146-184. 
1900b  Keller,  Arthur:  Phosphor  und  Stickstoff  im  Sauglingsorganismus,  Arch. 

Kinderheilk.  29,  1-95. 
1880    Kellner,  Oskar:  Versuche  iiber  die  Entbitterung  und  Verdaulichkeit  der 

Lupinenkorner,  Landwirtsch.  Jahrbticher  9,  977-998. 
1907     Kellner,   Oskar:   Die   Ernahrung  der     landwirtschaftlichen     Nutztiere, 

Berlin.      Fourth  edition.      621  pp. 
1894     Kellner,  O.,  A.  Kohler,  and  F.  Barnstein:  Untersuchungen  verschiedener 

Rauhfuttersarten  aus  Wirtschaften  in  denen  Knochenbriichigkeit  auf- 

tritt,  Sachs,  landwirtsch.  Zeit.  42,  167-171,  227. 


640  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1895  Kellner,  O.,  A.  Kohler  and  F.  Barnstein:  Untersuchungen  verschiedener 
Rauhfutterarten  aus  Wirtschaften,  in  denen  Knochenbriichigkeit  auf- 
tritt,  Biedermann's  Centralbl.  f.  Agriculturchem.  24,  441-443. 

1869a  Kemmerich,  E.:  Untersuchungen  uber  die  physiologische  Wirkung  der 
Fleischbruhe,  des  Fleischextracts  und  der  Kalisalze  des  Fleisches, 
Arch.  ges.  Physiol.  2,  49-93. 

1869b  Kemmerich,  E.:  Beitrage  zur  physiologischen  Chemie  der  Milch,  Ibid.  2, 
401-414. 

1909  Kennaway,  E.  L.,  and  J.  B.  Leathes:  A  Preliminary  Note  on  the  Examin- 

ation of  the  Fat  in  the  Liver  in  Health  and  Disease,  Proc.  Roy.  Soc. 
Med.  2,  Part  III,  Path.  Sect.,  136-144. 

1910  Kepinow,  L.:  Ueber  den  Einfluss  der  Blutkorperchenlipoide  auf  die  Blut- 

bildung,  Biochem.  Zeit.  30,  160-171. 

1908  Khuen,  A.:  Beitrage  zur  Kenntnis  des  Mineralstoffwechsels,  insbesondere 

des  Kalkes  beim  Wiederkauer,  Diss.,  Bonn.  41  pp. 

1909  Kida,  Y. :  Ueber  den  Einfluss  der  hoheren  Temperatur  beim  Sterilisieren 

der  Milch,  Jour,  of  College  of  Agr.,  Imperial  Univ.  Tokyo  1,  141-144. 
1907a  Kikkoji,  T.:  Ueber  das  Vorkommen  von  einem  Nucleinsaure  spaltenden 

Fermente  in  Cortinellus  edodes,  Zeit.  physiol.  Chem.  51,  201-206. 
1907b  Kikkoji,  T.:  Ueber  die  Nucleinsaure  aus  der  menschlichen  Placenta,  Ibid. 

53,  411-414. 

1909  Kikkoji,  T.:  Beitrage  zur  Kenntniss  des  Caseins  und  Paracaseins,  Ibid. 

61,  139-146. 

1910  Kilbourne,  E.  D.:  Food  Salts  in  Relation  to  Beriberi,  Philippine  Jour. 

of  Sci.,  B,  5,  127-135. 

1911  Kimura,  K.,  and  W.  Stepp:  Untersuchungen  iiber  den  Gehalt  des  Blut- 

serums  an  atherloslichem   Phosphor  bei  verschiedenen   Krankheiten, 

Deut.  Arch.  klin.  Med.  104,  209-215. 
1911     Kinberg,  Georg:  Beitrag  zur  Kenntnis  des  Stoffwechsels  bei  N- Hunger, 

Skand.  Arch.  Physiol.  25,  291-314. 
1906    Kitagawa,  F.,  and  H.  Thierf elder:  Ueber  das  Cerebron.      III.  Mitteilung, 

Zeit.  physiol.  Chem.  49,  286-292. 
1896     Klautsch,  A.:  Einige  Mittheilungen  iiber  die  Verwendbarkeit  von  H.  O. 

Opel's  Nahrzwieback  als  Nebenkost  fur  Sauglinge  und  an  Rachitis 

leidende  Kinder,  Jahrb.  Kinderheilk.  43,  190-202. 
1908     Kleinertz:  Die  Anwendung  von  physiologisch  reinem  Lecithin  in  der  Ther- 

apie,  Med.  Klinik  4,  191,  192. 
1899     Klemperer,  Georg:  Zur  Behandlung  der     Phosphaturie,     Therapie     der 

Gegenwart,  N.  F.  1,  351-354. 
1908     Klemperer,  Georg:  Ueber  Phosphaturie,  ein  Beitrag  zur  Prophylaxe  der 

Nierensteine,  Ibid.  49  (N.  F.  10),  3-11. 
1902     Klemperer,  G.,  and  F.  Tritschler:  Untersuchungen  iiber  Herkunft  und 

Loslichkeit  der  im  Urin  ausgeschiedenen  Oxalsaure,  Zeit.  klin.  Med. 

44,  337-366. 
1908    Klien,  G.:  Ueber  den  Wert  des  Beif utters    von    phosphorsaurem    und 

kohlensaurem   Kalk  bei  verschiedenen   Tiergattungen,   Milch-Zeitung 

1908,  No.  3. 
(1911)  Klieneberger,  Otto  L.:  Die  Behandlung  der  progressiven  Paralyse  mit 

Natrium  nucleinicum,  Berlin,  klin.  Wochenschr.  48,  330-333;  through 
Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  41  (1911)..  923. 


PHOSPHORUS  METABOLISM  641 

1906  Klimmer  and  Schmidt:  Beitrag  zur  Aetiologie  der  Halisteresis  ossiura 
nebst  therapeutischen  Bemerkungen,  Monatshefte  prakt.  Tierheilk.  17, 
481-517.      127  refs. 

1882    Klinkenberg,  W.:  Ueber  die  Nucleiine,  Zeit.  physiol.  Chem.  6,  566-571. 

1909     Klotz,  M.:  Milchsaure  und  Sauglingstoffwechsel,  Jahrb.  Kinderheilk.  70, 

1-61. 
1905    Klotz,  Oskar:  On  the  Presence  of  Soaps  in  the  Organism  in  Certain 

Pathological  Conditions  (A  Preliminary  Communication),  Proc.  Amer. 

Physiol.  Soc,  in  Amer.  Jour.  Physiol.  13,  XXI,  XXII. 
1893     Klug,  Ferd.,  and  Viktor  Olsavszky:  Einfluss  der  Muskelsarbeit  auf  die 

Ausscheidung  der  Phosphorsaure,  Arch.  ges.  Physiol.  54,  21-26. 
1908     Knapp,  Paul:  Experimenteller  Beitrag  zur  Ernahrung  von  Ratten  mit 

kiinstlicher  Nahrung  und     zum     Zusammenhang     von     Ernahrungs- 

storungen  mit  Erkrankungen  der  Conjunctiva,  Zeit.  exp.  Path.  u.  Ther. 

5,  147-169. 

1898  Knauthe,  Karl:  Zur  Kenntniss  des  Stoffwechsels  der  Fische,  Arch.  ges. 

Physiol.  73,  490-500. 
1898a  Knopf  elmacher,  Wilhelm:  Verdauungsriickstande  bei  der  Ernahrung  mit 

Kuhmilch  und  ihre  Bedeutung  fur   den   Saugling,   Beitrage   z.   klin. 

Med.  u.  Chir.  1898,  18,  7-75. 
1898b  Knopf  elmacher,  Wilhelm:  Ueber  Caseinverdauung,  Wien.  klin.  Wochen- 

schr.  11,  1024,  1025. 

1899  Knopf  elmacher,  Wilhelm:  Neue  Versuche  uber  Caseinausnutzung,  Ibid. 

12,  1308-1310. 

1900  Knopf  elmacher,  Wilhelm:  Versuche  iiber  die  Ausnutzung  des  Kuhmilch- 

kaseins,  Jahrb.  Kinderheilk.  52,  545-571. 
1914     Knopf,  Martin:  Ueber  das  Nucleoproteid  nach  Hammarsten  aus  Rinder- 

pankreas.      I.  Mitteilung,  Zeit.  physiol.  Chem.  89,  170-174. 
1888     Kobler,  G.:  Zur  Kenntniss  der  Osteomalacie,  Wien.  klin.  Wochenschr.  1, 

459-461;  482,  483. 
1900     Kobrak,  Erwin:  Beitrage  zur  Kenntniss  des  Caseins  der  Frauenmilch, 

Arch.  ges.  Physiol.  80,  69-85. 
1906     Koch,  Egbert:  Ein  Beitrag  zum  Phosphorstoffwechsel,  St.  Petersburger 

med.  Wochenschr.  31,  400-402. 
1913    Koch,  Mathilde  L.:  Contributions  to  the  Chemical  Differentiation  of  the 

Central  Nervous  System.      I.  A  Comparison  of  the  Brain  of  the  Albino 

Rat  at  Birth  with  That  of  the  Fetal  Pig,  Jour.  Biol.  Chem.  14,  267-279. 
1902a  Koch,  Waldemar:  The  Lecithans,  their  Function  in  the  Life  of  the  Cell, 

Univ.  of  Chicago  Decennial  Pub.  10,  1-12. 
1902b  Koch,  Waldemar:  Zur  Kenntniss  des  Lecithins,  Kephalins  und  Cerebrins 

aus  Nervensubstanz,  Zeit.  physiol.  Chem.  36,  134-140. 

1903  Koch,  Waldemar:  Die  Lecithane  und  ihre  Bedeutung  fur  die  lebende 

Zelle,  Ibid.  37,  181-188. 

1904  Koch,  Waldemar:  Methods  for  the  Quantitative  Chemical  Analysis  of 

the  Brain  and  Cord,  Amer.  Jour.  Physiol.  11,  303-329. 
1905a  Koch,  Waldemar:  Therapeutic  Value  of  Lecithin  in  Infant  Feeding,  St. 

Louis  Courier  of  Med.,  June  1905. 
1905b  Koch,  Waldemar:  Relation  of  Kreatinin  Excretion  to  Variations  in  Diet, 

Amer.  Jour.  Physiol.  15,  15-29. 


642  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1906    Koch,   Waldemar:  Ueber  der  Lecithingehalt  der  Milch,  Zeit.  physiol. 

Chem.  47,  327-330. 
1907a  Koch,  Waldemar:  The  Relation  of  Electrolytes  to  Lecithin  and  Kephalin, 

Jour.  Biol.  Chem.  3,  53-56. 
1907b  Koch,  Waldemar:  Zur  Kenntnis  der  Schwefelverbindungen  des  Nerven- 

systems,  Zeit.  physiol.  Chem.  53,  496-507. 
1909a  Koch,  Waldemar:  Phosphorus  Compounds  as  Brain  Foods,  Jour.  Amer. 

Med.  Assoc.  52,  1381-1383. 
1909b  Koch,  Waldemar:  Die  Bedeutung  der  Phosphatide  (Lecithane)  fur  die 

lebende  Zelle.      II.  Mitteilung,  Zeit.  physiol.  Chem.  63,  432-  442. 
1910a  Koch,  Waldemar:  Zur  Kenntnis  der  Schwefelverbindungen  des  Nerven- 

systems.      II.  Mitteilung.      Ueber  ein  Sulfatid  aus  Nervensubstanz, 

Ibid.  70,  94-97. 
1910b  Koch,  Waldemar:  Pharmacological     Studies  on    the    Phosphatids.      I. 

Methods  for  the  Study  of  their  Combinations  with  Drugs  and  Other 

Substances,  Jour.  Pharmacol,  and  Exp.  Therapeut.  2,  239-244. 

1912  Koch,  Waldemar:  Should  the  Term  Protagon  be  Retained,  Proc.  Soc 

Bi-ol.  Chem.,  in  Jour.  Biol.  Chem.  11,  XL. 

1906  Koch,  Waldemar,  and  William  H.  Goodson:  A  Preliminary  Study  of  the 

Chemistry  of  Nerve  Tissue  Degeneration,  Amer.  Jour.  Physiol.  15, 
272-279. 

1913  Koch,  W.,  and  M.  L.  Koch:  Contributions  to  the  Chemical  Differentiation 

of  the  Central  Nervous  System.  III.  The  Chemical  Differentiation  of 
the  Brain  of  the  Albino  Rat  during  Growth,  Jour.  Biol.  Chem.  15,  423- 
448. 
1910  Koch,  W.,  and  F.  C.  McLean:  Pharmacological  Studies  on  the  Phos- 
phatids. 3.  The  Relation  of  the  Phosphatids  to  Overton  and  Meyer's 
Theory  of  Narcosis,  Jour.  Pharmacol,  and  Exp.  Therapeut.  2,  249-252. 

1907  Koch,  Waldemar,  and  Sydney  A.  Mann:  A  Comparison  of  the  Chemical 

Composition  of  Three  Human  Brains  at  Different  Ages,  Proc.  Physiol. 
Soc,  in  Jour,  of  Physiol.  36,  XXXVI-XXXVIII. 

1909  Koch,  Waldemar,  and  Sydney  A.  Mann:  A  Chemical  Study  of  the  Brain 

in  Healthy  and  Diseased  Conditions,  with  Especial  Reference  to 
Dementia  Praecox,  Archives  of  Neurol,  and  Psychiatry  4,  2-46. 

1910  Koch,  W.,  and  H.  T.  Mostrom:  Pharmacological  Studies  on  the  Phos- 

phatids. 5.  The  Function  of  the  Brain  Phosphatids  in  the  Physiologi- 
cal Action  of  Strychnine,  Jour.  Pharmacol,  and  Exp.  Therapeut.  2, 
265-269. 

1910  Koch,  W.,  and  F.  H.  Pike:  Pharmacological  Studies  on  the  Phosphatids. 
2.  The  Relation  of  the  Phosphatids  to  the  Sodium  and  Potassium  of 
the  Neuron,  Ibid.  2,  245-248. 

1907  Koch,  W.,  and  Howard  S.  Reed:  The  Relation  of  Extractives  to  Protein 
Phosphorus  in  Aspergillus  Niger,  Jour.  Biol.  Chem.  3,  49-52. 

1910  Koch,  Waldemar,  and  W.  W.  Williams:  Pharmacological  Studies  on  the 
Phosphatids.  4.  The  Relation  of  Brain  Phosphatids  to  Tissue  Metab- 
olites, Jour.  Pharmacol,  and  Exp.  Therapeut.  2,  253-264. 

1905  Koch,  Waldemar,  and  Herbert  S.  Woods:  The  Quantitative  Estimation 
of  the  Lecithins,  Jour.  Biol.  Chem.  1,  203-211. 

1910  Kochmann,  Martin:  Zur  Kenntnis  des  Kalkstoffwechsels  unter  Berttck- 
sichtigung  des  Stoffwechsels  der  Phosphorsaure  und  der  Magnesia. 
Vorlaufige  Mitteilung,  Biochem.  Zeit.  27,  85,  86. 


PHOSPHORUS  METABOLISM  643 

1911  Kochmann,  Martin:  Ueber  die  Abhangigkeit  des  Kalkstoffwechsels  von 

den  organischen  Nahrungskomponenten  beim  erwachsenen  Hunde, 
nebst  Bemerkungen  iiber  den  Stoffumsatz  der  Phosphorsaure  und  der 
Magnesia.      I.  Mitteilung,  Ibid.  31,  361-376. 

1912  Kochmann,  Martin:  Zur  Wirkung  des  Phosphors  auf  den  Kalkstoffwech- 

sel  des  Hundes,  Ibid  39,  81-87. 

1911  Kochmann,  Martin,  and  Ernst  Petzsch:  Ueber  die  Abhangigkeit  des  Kalk- 
stoffwechsels von  den  organischen  Nahrungskomponenten  beim 
erwachsenen  Hunde  nebst  Bemerkungen  iiber  den  Stoffwechsel  der 
Phosphorsaure  und  der  Magnesia.  II.  &  III.  Mitteilungen,  Ibid  32, 
10-26;  27-42. 

1904  Kbhler,  A.,  F.  Honcamp,  M.  Just,  J.  Volhard,  M.  Popp,  and  0.  Zahn: 
Ueber  die  Assimilation  des  Kalkes  und  der  Phosphorsaure  aus  ver- 
schiedenen  Kalkphosphaten  durch  wachsende  Tiere,  Verhandl.  d.  76. 
Versamml.  d.  Gesell.  deut.  Naturforsch.  u.  Aerzte,  1904,  II,  pt.  1,  152- 
154;  also  Landwirtsch.  Versuch.  Stat.  61,  458-479. 

1907  Kohler,  A.,  F.  Honcamp,  and  P.  Eisenkolbe:  Weitere  Untersuchungen 
iiber  die  Assimilation  der  Phosphorsaure  und  des  Kalkes  aus  Kalkphos- 
phaten durch  wachsenden  Tiere,  Landwirtsch.  Versuch.  Stat.  65,  349- 
380. 

1903  Konig,  J.:  Chemie  der  menschlichen  Nahrungs-  und  Genussmittel.  4th 

Edition.  Vol.  I  (1903);  Chemische  Zusammensetzung  der  mensch- 
lichen Nahrungs-  und  Genussmittel.  1535  pp.  (By  A.  Bomer).  Vol. 
II  (1904) :  Die  menschlichen  Nahrungs-  und  Genussmittel,  ihre 
Herstellung,  Zusammensetzung  und  Beschaffenheit,  nebst  einem  Abriss 
iiber  die  Ernahrungslehre.  1557  pp.  Vol.  Ill,  pt.  1  (1910);  Unter- 
suchung  von  Nahrungs-,  Genussmitteln  und  Gebrauchsgegenstanden. 
1.  Teil.  Allgemeine  Untersuchungsverfahren.  772  pp.  2.  Teil. 
(Not  out  in  1913):  Die  Untersuchung  und  Beurteilung  der  einzelnen 
Nahrungs-,  Genussmittel  und  Gebrauchsgegenstande. 

1913a  Konig,  J.,  and  J.  Grossfeld:  Das  Fischsperma  als  Nahrungsmittel  fur 
den  Menschen,  Biochem.  Zeit.  54,  333-350. 

1913b  Konig,  J.,  and  J.  Grossfeld:  Der  Fischrogen  als  Nahrungsmittel  fur  den 
Menschen,  Ibid.  54,  351-394. 

1911  Koeppe,  Hans:  Studien  zum  Mineralstoffwechsel,  Jahrb.  Kinderheilk. 
73,  9-49. 

1904  Kovesi,  Geza,  and  Wilhelm  Roth-Schulz:  Pathologie  und  Therapie     der' 
Niereninsuffizienz  bei  Nephritiden  auf  Grund  eigener  Untersuchungen, 

Leipzig.       284  pp. 

1888  Kolpakcha,  J.  M.:  (On  the  Source  of  N  in  the  Urine),  Phiziologicheskii 
Sbornik,  Kharkov  1888,  Vol  1,  p.  53;  through  U.  S.  Dept.  Agr.,  Office 
Exp.  Sta.,  Bui.  45,  revised  edition  1898,  pp.  320-325. 

1910  Kondo,  Kura:  Ueber  die  Ausscheidung  von  organisch  gebundenem,  Phos- 
phor im  Ham,  Biochem.  Zeit.  28,  200-207. 

1902  von  Korczyriski,  L.  R.:  Zur  Kenntnis  des  Stoffwechsels  bei  Osteomalacic, 
Wien.  med.  Presse  43,  1073-1078;  1131-1133;  1177-1180;  1228-1232. 

1900  Kornauth,  Karl:   Fiitterungsversuche  mit   einem   Hunde     bei     Verab- 

reichung  verschiedener  stickstoffhaltiger  Materialien,  Zeit.  f.  d.  Land- 
wirtsch. Versuchswesen  in  Oesterreich  3,  1-25;  133-162. 

1901  Kornauth,  Karl:  Ueber  Fersan,  ein  neues  eisen-  und  phosphorhaltiges 

Nahrpraparat,  Zeit.  diatet.  u.  physikal.  Ther.  4,  480-486. 


644  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1900    Kornauth,  Karl,  and  O.  von  Czadek:  Ueber  ein  neues  Nahrpraparat 

"Fersan,"  Zeit.  f.  d.  Landwirthsch.  Versuchswesen  in  Oesterreich  3, 

556-573. 
1892    Kornblum,  Herwarth:  Ueber  die  Ausscheidung  des  Stickstoffs  bei  Nieren- 

krankheiten  des  Menschen  im  Verhaltniss  zur  Aufnahme  desselben, 

Arch.  path.  Anat.  u.  Physiol.  127,  409-445. 
1904    Kornf  eld,  F. :  Ueber  Protylin  und  seine  therapeutische  Verwertung,  Wien. 

med.  Presse  45,  2297-2304. 

1892  Korsakov,  N. :  Sur  la  reproduction  artificielle  du  rachitisme  chez  quelques 

animaux,  Internat.  zool.  kongress,  Moskau  1892,  261-267. 

1899  Kort,  A.:  (Ueber  den  Einfluss  der  Trachtigkeit  der  Kuh  auf  den  Gehalt 

der  Milch  an  Mineralstoffen,  insbesondere  an  Phosphorsaure  und  Kalk), 
L'Ingenieur  agricole  de  Gembloux  1,  453-475;  through  Jahresb.  ii.  d. 
Fortschr.  d.  Thierchem.  30  (1900),  256,  257. 

1879  Kossel,  Albrecht:  Ueber  das  Nuclei'n  der  Hefe,  Zeit.  physiol.  Chem.  3, 

284-291. 

1880  Kossel,  Albrecht:  Ueber  das  Nuclein  der  Hefe.      II.  Ibid.  4,  290-295. 

1881  Kossel,  Albrecht:  Untersuchungen  iiber  die  Nucleine  und  ihre  Spaltungs- 

producte,  Strasbourg.       19  pp. 

1882  Kossel,  Albrecht:  Zur  Chemie  des  Zellkerns,  Zeit.  physiol.  Chem.  7,  7-22. 

1885  Kossel,  Albrecht:  Ueber  das  Nuclei'n  im  Dotter  des  Hiihnereies,  Arch. 

Anat.  u.  physiol.  346,  347. 

1886  Kossel,   Albrecht:  Weitere   Beitrage   zur   Chemie   des   Zellkerns,   Zeit. 

physiol.  Chem.  10,  248-264. 
1891a  Kossel,   Albrecht,:   Ueber   die   chemische   Zusammensetzung   der   Zelle, 

Arch.  Anat.  u.  Physiol.,  physiol.  Abt.,  181-186. 
1891b  Kossel,  Albrecht:  Ueber  einige  Bestandtheile  des  Nervenmarks,  Ibid., 

359-366. 

1893  "Kossel,  Albrecht:  Ueber  die  Nucleinsaure,  Ibid.  157-164. 

1894  Kossel,  Albrecht :  Weitere  Beitrage  zur  Kenntniss  der  Nucleinsaure,  Ibid. 

194-200. 

1895  Kossel,  Albrecht:  Ueber  die  Aufnahme  des  Phosphors  in  den  tierischen 

Organismus,  Monatsschr.  Geburtsh.  u.  Gynakol.  1,  175-178. 

1896  Kossel,  Albrecht:  Ueber  die  Bildung  von  Thymin  aus  Fischsperma,  Zeit. 

physiol.  Chem.  22,  188-190. 

1900  Kossel,  Albrecht:  Bemerkungen  zu  der  Erwiderung  des  Herrn  Bang, 

Ibid.  31,  410. 

1911  Kossel,  Albrecht:  Chemische  Beschaffenheit  des  Zellkerns,  Munch,  med. 
Wochenschr.  58,  65-120. 

1893  Kossel,  A.,  and  Fr.  Freytag:  Ueber  einige  Bestandtheile  des  Nerven- 
marks und  ihre  Verbreitung  in  den  Geweben  des  Thierkorpers,  Zeit. 
physiol.  Chem.  17,  431-456. 

1893  Kossel,  A.,  and  Albert  Neumann:  Ueber  das  Thymin,  ein  Spaltungs- 

product  der  Nucleinsaure,  Ber.  deut.  chem.  Gesell.  26,  III,  2753-2756. 

1894  Kossel,  A.,  and  Albert  Neumann:  Darstellung    und     Spaltungsprodukte 

der  Nucleinsaure.       (Adenylsaure),  Ibid  27,  II,  2215-2222. 
1896     Kossel,  A.,  and  Albert  Neumann:  Ueber  Nucleinsaure  und  Thyminsaure, 

Zeit.  physiol.  Chem.  22,  74-81. 
1894     Kossel,  H.:  Ueber  die  Einwirkung  der  Nucleinsaure  auf  Bakterien,  Arch. 

Anat.  u.  Physiol.,  physiol.  Abt.  200-203. 


PHOSPHORUS  METABOLISM  645 

1903    Kostytschew,   S.:  Ueber  Thymonucleinsaure,  Zeit.   physiol.   Chem.  39, 

545-560. 
1912    Kovaliova,  M.  M.:  Influence  des     preparations    phosphorees     sur    les 

processus  d'oxydation  evoluant  dans  l'organisme  animal,  Archives  des 

sci.  biol.  17,  279-307. 
1910    Kowalevsky,  Katharina:  Ueber  die  Zusammensetzung  der  Nucleinsaure 

aus  Hefe,  Zeit.  physiol.  Chem.  69,  240-264. 
1903-4  von  Koziczkowsky,  Eugen :  Beitrage  zur  Kenntniss  des  salzstoffwechsels 

mit  besonderer  Beriicksichtigung  der  chronischen  Nephritiden,  Zeit. 

klin.  Med.  51,  287-330. 
1857    Krabbe,  H. :  Om  Phosphorsyremaengden  i  Urinen  og  om  de  Phosphorsure 

Jordarters   Udfaeldning   deraf  ved  kogning    (Ueber   die   Menge   der 

Phosphorsaure  im  Harn  und  iiber  die  Ausscheidung  der  Erdphosphate 

beim  Kochen  des  Harns),  Kjobenhavn  1857;  through  Arch.  path.  Anat. 

u.  Physiol.  11  (1857),  478-480. 

1907  Kramer,  Reinhard:  Ueber  die  Wirkung    des    phosphorsauren    und    des 

kohlensauren  Kalkes  als  Beigabe  bei  Verabreichung  von  Riibenblattem 

an  Milchkiihe,  Inaug.  Diss.,  Leipzig.       69  pp. 
1900    Krchivetz,  M.  J.  W.:  De  l'elimination  d'azote  et  d'acide  phosphorique  au 

cours  de  l'inanition  complete  soit  simple,  soit  accompagnee  d'excitation 

douloureuse,  Archives  des  sci.  biol.  8,  37-56. 
1893     Krehl,  L.:  Ueber  die  fettige  Degeneration  des  Herzens,  Deut.  Arch.  klin. 

Med.  51,  416-450;  through  Jahresb.  u.  d.  Fortschr.  d.  Thierchem.  23 

(1893),  371-373. 

1895  Kriiger,  Friedrich:  Ueber  den  Schwefel-  und  Phosphorgehalt  der  Leber- 

und  Milzzellen  in  verschiedenen  Lebensaltern,  Zeit.  Biol.  31,  400-412. 

1896  Kriiger,  Th.  Richard:  Ueber  die  Abspaltung  von  Kohlensaure  aus  Phos- 

phorfleischsaure  durch  Hydrolyse,  Zeit.  physiol.  Chem.  22,  95-102. 
1899  Kriiger,  Th.  Richard:  Zur  Kenntniss  der  Nucleone,  Ibid.  28,  530-534. 
1895     Kiihnau,  W.:  Experimentelle  und  klinische  Untersuchungen     iiber     das 

Verhaltniss  der  Harnsaureausscheidung  zu  der  Leukocytose,  Zeit.  klin. 

Med.  28,  534-566. 
1896-7  Kiihnau,  W.:  Ueber  das  Verhalten  des  Stoffwechsels  und  der  "weissen 

Blutelemente  bei  Blutdissolution,  Deut.  Arch.  klin.  Med.  58,  339-367. 
1895    Kiihns,  C:  Untersuchungen  iiber  die  chemische  Zusammensetzung  der 

harten  Zahnsubstanzen  des  Menschen  in  verschiedenen  Altersstufen, 

Deut.  Monatsschr.  Zahnheilk.  13,  361-377;  450-465. 

1908  Kiinzel,  Werner,  and  Alfred  Schittenhelm :  Zur  Frage  des  Nucleinstoff- 

wechsels  beim  Menschen.      Vorl.  Mitt.,  Zentralbl.  ges.  Physiol,  u.  Pathc 
des  Stoffwechsels  N.  F.  3,  721-724. 

1909  Kiinzel,  Werner,  and  Alfred  Schittenhelm:  Gegenseitige  Beeinflussung 

der  Fermente  des  Nucleinstoffwechsels,  Zeit.  exp.  Phys.  u.  Ther.  5, 
393-400. 
1907    Kiittner,  S.:  Ueber  den  Einfluss  des  Lecithins  auf  die  Wirkung  der 
Verdauungsfermente,  Zeit.  physiol.  Chem.  50,  472-496. 

1909  Kiittner,   S.:  Peptische  Verdauung  des  Kaseins  vom   Standpunkte  der 

Aciditat  seiner  Spaltungsprodukte,  Arch.  ges.  Physiol.  129,  557-602. 

1910  Kutanin,  Michael:  Beitrage  zur  Kenntnis  der  chemischen  Zusammenset- 

zung des  Gehirns  bei  verschiedenen  Tieren,  Inaug.  Diss.,  Berlin,  1910; 
through  Jahresb.  u.  d.  Fortschr.  d.  Thierchem.  40  (1910),  445. 


646  OHIO  EXPEEIMENT  STATION:  TECHNICAL  BUL.  5 

1901     Kutscher,  Fr.:  Das  proteolytische  Enzym  der  Thymus.      I.  Mittheilung,, 

Zeit.  physiol.  Chem.  34,  114-118. 
1903     Kutscher,  Fr.,  and  Lohmann:  Die  Endprodukte  der  Pankreas-  und  Hefe- 

selbstverdauung,  Ibid.  39,  159-164;  313-317. 
1906     Kutscher,  Fr.,  and  Lohmann:  Der  Nachweis  toxischer  Basen  im  Harn, 

Ibid.  48,  1-8. 

1903  Labbe,  Henri:  La  lecithine,  Revue  de  therapeut.  70,  721-732. 

1904  Labbe,   Henri:   Les   medications   reconstituentes.    La   medication   phos- 

phoree, Paris.       96  pp. 
1890     Laehr,  H. :  Versuche  iiber  den  Einfluss  des  Schlaf es  auf  den  Stoffwechsel, 

Allgem.  Zeit.  Psychiat.  46,  286-317. 
1894     Lafevre:  Nombreux  cas  de  cachexie  ossifrage  observes  chez  les  vaches 

laitieres  par  suite  de  le  grande  secheresse,  Annales  de  med.  veterin. 

43,  504-510. 

1905  Laguesse:  Medication  phosphoree.     Essais  experimentaux  et  chimiques 

d'une  albumine  phosphoree  synthetique,  These  de  Paris.       88  pp. 
1884     Lailler,  A.:  Sur  l'elimination  de  l'acide  phosphorique  par  l'urine  dans 

l'alienation  mentale  et  l'epilepsie,  Compt.  rend.  Acad,  des  sci.  99,  572, 

573. 
1901     Lancereaux  and  Paulesco :  Note  sur  l'emploi  therapeutique  de  la  lecithine, 

Bui.  Acad,  de  med.  de  Paris  45,  685-688. 
1894     Landauer,  Armin-tol:      Adatok    A    Viz     Szerepehez    A    Szervezetben. 

(Beitrage  zur  Rolle  des  Wassers  im  Organismus),  Mathematikai  es 

termeszettudomanyi  ertesito,  Budapest,  1894,    12,    197;    also    Ungar. 

Arch.  Med.  3,  136-188;  through  Jahresb.  ii.  d.  Fortschr.  d.  Thierchem. 

24   (1894),  532-535. 
1912     Landsberg,  E.:  Untersuchungen  iiber  den   Stoffwechsel  von  Stickstoff, 

Phosphor  und  Schwefel     bei     Schwangeren,  Z.     Geburtsh.     71,  163; 

through  Chem.  Abs.  7  (1913),  2413. 

1906  Landsberg,  G. :  Das  Lecithin,  seine  Rolle  in  Organismus  und  seine  thera- 

peutische   Verwendung,   Zentralbl.   ges.   Physiol,   u.   Path,   des   Stoff- 

wechsels  7  (N.  F.  1),  193-212. 
1892     Landsteiner,  Karl:  Ueber  den  Einfluss  der  Nahrung  auf  die  Zusammen- 

setzung  der  Blutasche,  Zeit.  physiol.  Chem.  16,  13-19. 
1906     Lane,  A.  H.:  Bone  Diseases  Amongst  Horses  in  South  Africa,  Veterin. 

Jour.  62  (n.  s.  13),  232-245. 
1897     de  Lange,  C.  C:  Vergelijkende  asch-analyses,  Diss.,  Amsterdam  1897; 

through  Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  27  (1897),  260-265. 
1900     de  Lange,  Cornelia:  Die  Zusammensetzung  der  Asche  des  Neugeborenen 

und  der  Muttermilch,  Zeit.  Biol.  40  (N.  F.  22),  526-528. 
1903     de   Lange,   Cornelia:   Zur   Casuistik  der   Phosphaturie   im   Kindesalter, 

Jahrb.  Kinderheilk.  57,  93-95. 
1877     Langendorff,  O.,  and  J.  Mommsen:  Beitrage  zur  Kenntniss  der  Osteo- 
malacic, Arch.  path.  Anat.  u.  Physiol.  69,  452-487. 
1875     Langgaard,   Alexander:   Vergleichende   Untersuchungen   iiber   Frauen-, 

Kuh-  und  Stutenmilch,  Ibid  65,  1-9. 
1910     Langheld,  K.:  Ueber  Metaphosphorsaure-athylester  und  dessen  Anwend- 

ung  in  der  organischen  Chemie,  Ber.  deut.  chem.  Gesell.  43,  II,  1857- 

1860. 


PHOSPHORUS  METABOLISM  647 

• 

1911  Landheld,  K.:  Ueber  Ester  und  Amide  der  Phosphorsauren.  II.  Ueber 

Versuche  zur  Darstellung  den  Lecithinen  verwandter  Korper,  Ibid. 
.  44,  II,  2076-2087. 

1912  Langheld,  K.:  Ueber  Ester  und  Amide  der  Phosphorsaure.       III.  Ueber 

dioxyaceton-  und  Fructose-phosphorsaure,  Ibid.  45,  1125-1127. 
1903     Langstein,  Leo:  Bemerkungen  uber  das  Ovomukoid,  Beitrage  z.  chem. 

physiol.  u.  Path.  3,  510-513. 
1906     Langstein,  Leo:  Zur  Klinik  der  Phosphaturie,  Med.  Klinik,  Berlin,  2,  406, 

407. 
1910     Langstein,  L.,  and  F.  Edelstein:  Ueber  die  Einheitlichkeit  des  Frauen- 

milchkaseins,  Jahrb.  Kinderheilk.       72   (Erganzungsheft),  1-15. 
1910     Langstein,   Leo,   and  Albert   Memann:   Ein  Beitrag  zur  Kenntnis   der 

Stoffwechselvorgange  in  den  ersten  vierzehn  Lebenstagen  normaler 

und  friihgeborener  Sauglinge,  Ibid.  71,  604-611. 

1913  Lanzoni,  O.:  Modificazioni  indotte  da  alcuni  medicamenti  sulla  percent- 

uale  degli  elementi  costitutivi  del  latte  di  vacca,  Clinica  veterinaria 

36,  11-23,  58-69. 
1910     Lapidus,  Herman:  Diastase  und  Handelslecithin,  Biochem.  Zeit.  30,  39-55. 
1902     Larned,  E.  R.:  Concerning  a  New  Series  of  Synthetic  Salts;  The  Nucleids 

of  Iron,  Copper,  Mercury  and  Silver,  Therapeutic  Gazette,  Sept.  15, 

1902;  through  Jour.  Amer.  Med.  Assoc.  39,  II,  (1902),  938. 
1905    Laumonier,  J.:  Remineralisation  phosphoree,  Bui.  gener.  de  therapeut. 

150,  779-789. 
1910     Laurer,  G. :  Ueben  die  Futterverhaltnisse  einen  Einfluss  auf  die  Knochen- 

starke  aus?       Deut.  landw.  Tierzucht  14,  437-439;  through  Exp.  Sta. 

Record  24  (1911),  175. 
1909     Lauxen,  Nikolaus:  Experimentelle  Untersuchungen  liber  die  Einwirkung 

des  Phosphors  auf  Knochen-  und  Blutbildung  bei  wachsenden  Hunden, 

Inaug.  Diss.,  Bonn.       40  pp. 
1900     Laves,  E.:  Ueber  das  Eiweissnahrmittel  "Roborat"  und  sein  Verhalten 

im  Organismus,  vergleichen  mit  ahnlichen  Praparaten,  Munch,  med. 

Wochenschr.  47,  1339-1345. 
1903a  Laves,  E.:  Ueber  Farbstoff,  Lecithin  und  Fett  des  Eidotters  (Vorlaufige 

Mitteilung),  Pharmazeut.  Zeitung  48,  814-816. 
1903b  Laves,  E.:  Ueber  Lezithin  und  seine  Anwendungsform,  Verhandl.  d.  75. 

Versamml.  d.  Gesell.  deut.  Naturforsch.  u.  Aerzte,  1903,  II,  86-88. 
1883     Lawes,  J.  B.,  and  J.  H.  Gilbert:  Composition  of  the  Ash  of  the  Entire 

Animals,  and  of  Certain  Separated  Parts,  Phil.  Trans,  of  the  Royal 

Soc.  174,  865-890. 
1908-9  Laxa,  Otakar :  La  production  et  les  qualites  du  lait  de  la  brebis  valaque, 

Revue  gener.  du  lait  7,  289-300;  313-328;  337-347;  361-373;  391-399. 
1909     Leach,  Albert  E.:  Food  Inspection  and  Analysis,  New  York. 

1900  Lebbin,  Georg:  Ueber  den  Verteilung  der  Nahrstoffe  in  den  Huhnereiern, 

Zeit.  offentliche  Chem.  6,  148,149. 

1901  Lebbin,  Georg:  Der  Nahrwert  der  Huhnereier,  Therapeut.  Monatsh.  15, 

552,  553. 
(1910a)  von  Lebedew,  A.:    (Investigations  on  the  Non-cellular  Fermentation 
Process  with  the  Help  of  the  Ultrafilter),  Biochem.  Zeit.  20,  114-125; 
through  Chem.  Abs.  4  (1910),  53,  54. 


648  OHIO  EXPEEIMENT  STATION:  TECHNICAL  BUL.  5 

1910b  von  Lebedew*  A.:  Ueber  Hexosephosphorsaureester.      I.  Ibid.  28,  213- 

229. 
1911a  von  Lebedew,  A.:  Ueber  Hexosephosphorsaureester.       II.  Ibid.  36,  248- 

260;  through  Zeit.  Unters.  d.  Nahrungs-  u.  Genussmittel  24  (1912), 

283. 
(1911b)  von    Lebedew,    A.:  (The    Mechanism    of    Alcoholic    Fermentation), 

Compt.  rend.  Acad,  des  sci.  153,  136-138;  through  Chem.  Abs.  5  (1911), 

3298. 
(1911c)  von  Lebedew,  A.:  Ueber  den  Mechanismus  der  alkoholischen  Garung, 

Ber.  deut.  chem.  Gesell.  44,  2932-2942;  Annal.  Inst.  Pasteur   25,  847- 

851;  through  Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  41  (1911),  781. 
1912    von  Lebedew,  A.,  and  N.  Griaznoff :  Ueber  den  Mechanismus  der  alkohol- 
ischen Garung.      II.  Ibid.  45,  3256-3272. 
1906     LeClerc,  J.  A.,  and  F.  C.  Cook:  Metabolism  Experiments  with  Organic 

and  Inorganic  Phosphorus,  Jour.  Biol.  Chem.  2,  203-216. 
1909     Le  Clerc,  J.  A.,  and  Robert  Wahl:  Chemical  Studies  of  American  Barleys 

and  Malts,  U.  S.  Dept.  Agr.,  Bur.  Chem.  Bui.  124.      75  pp. 
1911     Lederer,  Richard,  and  Karl  Stolte:  Die  Zusammensetzung  des  Menschen- 

und  des  Hund-herzens,  Biochem.  Zeit.  35,  108-112. 
1842     Lehmann,  C.  G.:  Untersuchimgen  iiber  den  menschlichen  Harn,  Jour.  f. 

prakt.  Chem.'  27,  257-274. 

1893  Lehmann,  Curt,  Friedrich  Miiller,  Immanuel  Munk,  H.  Senator,  and  N. 

Zuntz:  Untersuchungen  an  zwei  hungernden  Menschen,  Arch.  path. 
Anat.  u.  Physiol.  131  Suppl.  1-228. 
1882     Lehmann,  Ernst:  Zur  Wirkung  des  kohlensauren  Kalks  und  der  kohlen- 
sauren  Magnesia,  Berlin,  klin.  Wochenschr.  19,  320-324. 

1894  Lehmann,  Ernst:  Zur  Wirkung  des  kohlensauren  Kalkes,  Ibid.  31,  538- 

540. 

1859  Lehmann,  Jul.:  Ueber  die  mineralischen  Nahrstoffe,  insbesondere  iiber 
die  Erdphosphate  als  Nahrstoff  des  jungen  thierischen  Organismus, 
Landwirtsch.  Versuch.  Stat.  1,  68-86;  also  Annal.  der  Chem.  u.  Pharm. 
108  (1858),  357-378. 

1873  Lehmann,  Jul.:  Ueber  den  Werth  des  Fleischfuttermehls,  Zeit.  d.  land- 
wirtsch. Vereins  in  Bayern,  Dec.  1873;  through  Jahresb.  ii.  d.  Fortschr. 
d.  Agriculturchem.  17  (1876),  183,  184. 

1877  Lehmann,  Jul.:  Den  Einfluss  der  Nahrung  auf     die     Knochenbildung, 

Amtlicher  Ber.   d.    50.  Versamml.   deut.   Naturforsch.   u.   Aerzte   in 
Miinchen,  215. 

1878  Lehmus,  Emilie:  Ueber  den  relativen  Werth  der  Phosphorsaure  im  Urin 

beim  Kinde,  Central-Zeitung  Kinderheilk.  1,  291,  292. 
1903     Leichnam,  Charles:  Urologie  comparee  du  rachitisme  et  de  la  scoliose 

des  adolescents.  Theorie  rachitique  de  la  scoliose.     Traitement  general 

de  la  scoliose,  These  de  Paris.      49  pp. 
1899     Leipziger,  Richard:  Ueber  Stoffwechselversuche  mit  Edestin,  Arch.  ges. 

Physiol.  78,  402-422;  also  Inaug.  Diss.,  Breslau,  1899. 

1901  Leo,  Hans:  Zur  Phosphorbehandlung  der  Rachitis,  Verhandl.  d.  Versamml. 

d.  Gesell.  f.  Kinderheilk.  18,  145-148. 

1902  Leo,  Hans:  Ueber  Alkalinurie,  Deut.  Arch.  klin.  Med.  73,  604-615. 

1909  Leonard,  V.  N.,  and  Walter  Jones:  On  Preformed  Hypoxanthin,  Jour. 
Biol.  Chem.  6,  453-460. 


PHOSPHORUS  METABOLISM  649 

1901  Lepine,  R.:  Sur  la  relation  existant  entre  l'etat  graisseux  du  foie  (avec 

augmentation  de  la  proportion  de  la  lecithine  hepatique)   et  le  phos- 

phore  incompletement  oxyde  de  l'urine,  Compt.  rend.  Soc.  de  biol.  53, 

978,979. 
1882     Lepine,  R.,  and  Eymonnet:  Sur  la  determination  quantitative  de  l'acide 

phosphoglycerique  dans  l'urine,  a  l'etat  physiologique  et  dans  diverses 

conditions  anomales,  notamment  dans  le  cas  de  foie  gras,  Ibid.  34,  622- 

625. 
1884     Lepine,  R.:  Eymonnet,  and  Aubert:  Sur    la    proportion    de    phosphore 

incompletement  oxyde  contenue     dans     l'urine,     specialement     dans 

quelques  etats  nerveux,  Ibid.  1884,  499,  500;  also  Compt.  rend.  Acad. 

des  sci.  98  (1884),  238-241. 
1879     Lepine,  R.:  and  Jacquin:  Sur  l'excretion  de    l'acide    phosphorique    par 

l'urine  dans  ses  rapports  avec  celle  de  l'azote,  Revue  mensuelle  de  med. 

et  de  chir.  3,  449-456;  716-728;  959-968. 

1902  Lepine,  R.:  and  Maltet:  Sur  l'elimination  de  l'acide  phosphorique  dans  la 

glycosurie  experimental e,  Compt.  rend.  Soc.  de  biol.  54,  921,  922. 
1905     Lepski,  Chaim:  Phosphortherapie  der  Rachitis,  Diss.,  Berlin.  34  pp. 

1903  Lesem,  W.  W.,  and  William  J.  Gies:  Notes  on  the  "Protagon"  of  the  Brain, 

Amer.  Jour.  Physiol.  8,  183-196. 
1913     Leubuscher,  Paul :   Therapeutische  Versuche  mit   Phosphor  bei  Epilepti- 

kern,  Deut.  med.  Wochenschr.  39,  494-496. 
1899     Levene,  P.  A.:  On  the  Nucleoproteid  of  the  Brain,  Archives  of  Neurol. 

and  Psychopathol.  2,  3-14. 
1910a  Levene,  P.  A.:  Ueber  das  Ichthulin  des  Kabeljau,  Zeit.  physiol.  Chem.  32, 

281-284. 
1901b  Levene,  P.  A.:  Darstellung  und  Analyse  einiger  Nukleinsauren,  Ibid.  32, 

541-551. 
1903a  Levene,  P.  A.:  Darstellung  und  Analyse  einiger  Nukleinsauren.  (Zweite 

Mittheilung),  Ibid.  37,  402-406. 
1903b  Levene,  P.  A.:  Darstellung  und  Analyse  einiger  Nucleinsauren.  Dritte 

Mitteilung,  Ibid.  38,  80-83. 
1903c  Levene,  P.  A.:  On  the  Chemistry  of  the  Chromatin  Substance  of  the 

Nerve-Cell,  Jour,  of  Med.  Research  10  (N.  S.  5),  204-211. 
1903d  Levene,  P.  A.:  Darstellung  und  Analyse  einiger  Nucleinsauren.       (Vierte 

Mitteilung.)  Zeit.  physiol.  Chem.  39,  4-8. 
1903e  Levene,  P.  A.:  Darstellung  und  Analyse  einiger  Nucleinsauren.  (Funfte 

Mitteilung.)  Ibid.  39,  133-135. 
1903f    Levene,  P.  A.:  Darstellung  und  Analyse  einiger  Nucleinsauren.  (Sechste 

Mitteilung.)       Ibid.  39,  479-483. 

1904  Levene,  P.  A.:  Darstellung  und  Analyse  einiger  Nucleinsauren.       (Sie- 

bente  Mitteilung.)       Ibid.  43,  199-201. 

1905  Levene,  P.  A.:  Darstellung  und  Analyse  einiger  Nucleinsauren.       VIII. 

Mitteilung.  Ueber  die  Milznucleinsaure,  Ibid  45,  370-380. 
1909a  Levene,  P.  A.:  Ueber  die  gepaarten  Phosphorsauren  in  Pflanzensamen, 

Biochem.  Zeit.  16,  399-405. 
1909b  Levene,  P.  A.:  Ueber  die  Hefenucleihsaure,  Ibid.  17,  120-131. 
1910     Levene,  P.  A.:  On  the  Biochemistry  of  Nucleic  Acids,  Jour.  Amer.  Chem. 

Soc.  32,  231-240. 
1901     Levene,  P.  A.,  and  C.  Alsberg:  Zur  Chemie  der  Paranucleinsaure,  Zeit. 

physiol.  Chem.  31,  543-555. 


650  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1908  Levene,  P.  A.,  and  W.  A.  Jacobs:  Ueber  die  Inosinsaure.  I.  Mitt.  Ber. 
deut.  chem.  Gesell.  41,  II,  2703-2707. 

1909a  Levene,  P.  A.,  and  W.  A.  Jacobs :  Ueber  Inosinsaure.  II.  Mitt.  Ibid  42,  I, 
335-338. 

1909b  Levene,  P.  A.:  Ueber  Inosinsaure.      III.  Mitt.  Ibid.  42,  I,  1198-1203. 

1909c  Levene,  P.  A.,  and  W.  A.  Jacobs:  Ueber  die  Pentose  in  den  Nucleinsauren, 
Ibid.  42,  II,  2102-2106. 

1909d  Levene,  P.  A.,  and  W.  A.  Jacobs:  Ueber  die  Pentose  in  den  Nuclein- 
sauren.      II.  Mitt.  Ibid.  42,  III,  3247-3251. 

1909e  Levene,  P.  A.,  and  W.  A.  Jacobs:  Ueber  Guanylsaure.  (I.  Mitteilung.) 
Ibid.  42,  II,  2469-2473. 

1909f  Levene,  P.  A.,  and  W.  A.  Jacobs:  Ueber  die  Hefe-Nucleinsaure.  I.  Ibid. 
42,  II,  2474-2478. 

1909g  Levene,  P.  A.,  and  W.  A.  Jacobs:  Ueber  die  Hefe-Nucleinsaure.  II.  Ibid. 

42,  II,  2703-2706. 

1910     Levene,  P.  A.  and  W.  A.  Jacobs:  Ueber  die  Hefe-  Nucleinsaure.    III.  Ibid. 

43,  III,  3150-3163. 

1911a  Levene,  P.  A.,  and  W.  A.  Jacobs:  Ueber  die  Inosinsaure.  IV.  Ibid.  44, 
I,  746-753. 

1911b  Levene,  P.  A.,  and  W.  A.  Jacobs:  Ueber  die  Hefe-Nucleinsaure.  IV. 
Ibid.  44,  1027-1032. 

1912a  Levene,  P.  A.,  and  W.  A.  Jacobs:  Guaninehexoside  Obtained  by  Hydroly- 
sis of  Thymus  Nucleic  Acid,  Jour.  Biol.  Chem.  12,  377-379. 

1912b  Levene,  P.  A.,  and  W.  A.  Jacobs:  On  the  Structure  of  Thymus  Nucleic 
Acid,  Ibid.  12,  411-420. 

1912c  Levene,  P.  A.,  and  W.  A.  Jacobs :  On  Guanylic  Acid.  Second  paper.  Ibid. 
12,  421-426. 

1910  Levene,  P.  A.,  and  F.  B.  LaForge:  Ueber  die  Tritico-Nucleinsaure,  Ber. 
deut.  chem.  Gesell.  43,  III,  3164-3167. 

1912  Levene,  P.  A.,  and  W.  A.  Jacobs:  Ueber  die  Hefe-Nucleinsaure.      V.  Die 

Struktur  der  Pyrimidin-Nucleoside,  Ibid.  45,  I,  608-620. 

1913  Levene,  P.  A.,  and  W.  A.  Jacobs:  On  Nucleases.       Third  paper,  Jour. 

Biol.  Chem.  13,  507-509. 
1906a  Levene,  P.  A.,  and  John  A.  Mandel:  Ueber  die  Kohlehydratgruppe  des 

Milznucleoproteids,  I.  Mitt.  Zeit.  physiol.  Chem.  47,  151-153. 
1906b  Levene,  P.  A.,  and  John  A,  Mandel:  Darstellung  und  Analyse  einiger 

Nucleinsauren.  XI.  Ueber  die  Nuclei'nkorper  des  Eies  des  Schellfisches, 

Ibid.  49,  262-265. 
1906c  Levene,  P.  A.,  and  John  A.  Mandel:  Darstellung  und  Analyse  einiger 

Nucleinsauren.       XII.   Mitteilung.       Nucleinsaure  der   Spermatozoen 

des  Maifisches  (Alosa)  Ibid.  50,  1-9. 
1908a  Levene,  P.  A.,  and  John  A.  Mandel:  Ueber  die  Konstitution  der  Thymo- 

nucleinsaure,  Ber.  deut.  chem.  Gesell.  41,  II,  1905-1909. 
1908b  Levene,  P.  A.,  and  John  A.  Mandel:  Zur  Chemie  der  Lebernucleoproteide. 

I.  Ueber  die  Guanylsaure,  Biochem.  Zeit.  10,  221-228. 
1911a  Levene,  P.  A.,  and  F.  Medigreceanu :  Nuclein  Metabolism  in  the  Dog, 

Amer.  Jour.  Physiol.  27,  438-445. 
1911b  Levene,  P.  A.,  and  F.  Medigreceanu:  On  Nucleases,  Jour.  Biol.  Chem.  9, 

65-83. 
1911c  Levene,  P.  A.,  and  F.  Medigreceanu:  The  Action  of  Gastro-Intestinal 

Juices  on  Nucleic  Acids,  Ibid.  9,  375-387. 


PHOSPHORUS  METABOLISM  651 

1911d  Levene,  P.  A.,  and  F.  Medigreceanu :  On  Nucleases.      Second  paper.  Ibid. 
9,  389-402. 

1904  Levene,  P.  A.,  and  L.  B.  Stookey:  Notiz  iiber  das  Pankreasnucleoproteid, 

Zeit.  physiol.  Chem.  41,  404-406. 
1913     Levene,  P.  A.,  and  C.  J.  West:  The  Saturated  Fatty  Acid  of  Kephalin, 
Jour.  Biol.  Chem.  16,  419-422. 

1905  Levy,   Fritz:   Ueber  den  therapeutischen  Wert  des   Lecithins  und  der 

lecithinhaltigen  Nahrpraparate  (Lecitogen),  Berlin,  klin.  Wochenschr. 
42,  1242-1245. 
1894     Levy,  Moritz :  Chemische  Untersuchungen  iiber  osteomalacische  Knochen, 
Zeit.  physiol.  Chem.  19,  239-270. 

1905  Lewin,  Carl:  Ueber  das  Lecithin  und  Bromlecithin,  Med.  Klinik  (Berlin) 

1,  857,  858. 
1905-8  Lewite,  Maxim :  Zur  Frage  des  Zusammenhanges  zwischen  Wetter,  Min- 

eralstoffgehalt  der  Futterpflanzen  und  Knochenbruchigkeit  des  Rindes, 

Mitt.  d.  landwirtsch.  Inst,  in  Leipzig  8,  69-111. 
1911     Lewoniewska,  S.:  Schwankungen  in  dem  Gehalt  der  Pflanzensamen  an 

einzelnen  Phosphorsaureverbindungen     in     ihrer     Abhangigkeit     von 

Vegetationsbedingungen,  Anzeiger  Akad.  Wiss.  Krakau  1911,  Reihe 

B,  85-96;  through  Zeit.  Unters.  d.  Nahrungs-  u.  Genussmittel  26  (1913), 

252,  253. 
1894     Leze,  R.,  and  E.  Hilsont:  Essai  des  laits  par  la  presure,  Compt.  rend. 

Acad,  des  sci.  118,  1069-1071. 
1888a  Liebermann,  Leo:  Embryochemische  Untersuchungen.     1.  Ueber  einige 

weniger  bekannte  Bestandtheile  des  Huhnereies,  Arch.  ges.  Physiol.  43, 

71-151. 
1888b  Liebermann,  Leo:  Ueber  das  Nuclein  der  Hefe  und  kunstliche  Darstellung 

eines  Nucleins  aus  Eiweiss  und  Metaphosphorsaure,  Ber.  deut.  chem. 

Gesell.  21,  I,  598-600. 

1889  Liebermann,  Leo:  Ueber  das  Nuclein  der  Hefe  und  kunstliche  Darstell- 

ung eines  Nucleins  aus  Eiweiss  und  Metaphosphorsaure,   Centralbl. 
med.  Wissensch.  27,  210-212;  225-227;  447. 

1890  Liebermann,  Leo:  Nachweis  der  Metaphosphorsaure  im  Nuclein  der  Hefe, 

Arch.  ges.  Physiol.  47,  155-160. 
1891a  Liebermann,  Leo:  Studien  iiber  die  chemischen  Processe  in  der  Magen- 

schleimhaut,  Ibid.  50,  25-54. 
1891b  Liebermann,  Leo:  Notiz  iiber  das  chemische  Verhalten  des  Nierenparen- 

chyms,  Ibid.  50,  55,  56. 
1891c  Liebermann,  Leo:  Ueber  den  Phosphorsauregehalt  des  Pferdeharn  unter 

physiologischen  und  pathologischen  Verhaltnissen,  Ibid.  50,  57-65. 
1893a  Liebermann,  Leo:  Neuere  Untersuchungen  iiber  das  Lecithalbumin,  Ibid. 

54,  573-585. 
1893b  Liebermann,  Leo:  Studien  iiber  die  chemischen  Vorgange  bei  der  Ham- 
secretion,  Ibid.  54,  585-606. 
1893     Liebermann,  L.,  and  Belav  v.  Bitto:  Ueber  Nucleinsaure,  Centralbl.  med. 

Wissensch.  1893,  Nos.  28  and  45. 

1906  Liebermeister,  G.:  Ueber  das  Nukleoproteid  des  Blutserums,  Beitrage  z. 

chem.  Physiol,  u.  Path.  8,  439-444. 
1847     Liebig,  Justus:  Ueber  die  Bestandtheile  der  Fliissigkeiten  des  Fleisches, 
Annalen  der  Chem.  u.  Pharm.  62,  257-369. 


652  OHIO  EXPERIMENT  STATION:  TECHNICAL  BTJL.  5 

1851    Liebig,  Justus:  Letters  28  and  29,  Familiar  Letters  on  Chemistry,  Third 

edition. 
1865     Liebreich,  Oscar:  Ueber  die  chemische  Beschaffenheit  der  Gehirnsub- 

stanz,  Annalen  der  Chem.  u.  Pharm.  134,  29-44. 

1894  Lilienfeld,  Leon:  Zur  Chemie  der  Leucocyten,  Zeit.  physiol.  Chem.  18, 

473-486. 

1895  Lilienfeld,  Leon:  Ueber  Blutgerinnung,  Ibid.  20,  89-165. 

1893  Lilienfeld,  Leon,  and  Achille  Monti:  Ueber  die  mikrochemische  Local- 

isation des  Phosphors  in  den  Geweben,  Ibid.  17,  410-424. 

1894  von  Limbeck,  R.:  Zur  Kenntniss  der  Osteomalacic,  Wien.  med.  Woch- 

enschr.  44,  737-739;  793-796;  844-847. 
1898     Lindemann,  W. :  Zur  Toxikologie  der  organischen  Phosphorverbindungen, 

Arch.  exp.  Path.  u.  Pharm.  41,  191-217. 
1912a  Lindet,  L.:  Sur  les  elements  mineraux  contenus  dans  la  caseine  du  lait, 

Rep.  Eighth  Internat.  Congr.  of  Applied  Chem.  19,  199-207. 
1912b  Lindet,  L.:  Sur  les  formes  que  le  phosphore  et  le  calcium  affectent  dans 

la  caseine  du  lait,  Compt.  rend.  Acad,  des  sci.  155,  923,  924;  also  Bui. 

Soc.  chim.  11  (1912),  950-952. 
1913a  Lindet,  L.:  Solubilite  des  albuminoiides  du  lait  dans  les  elements  du 

serum;  retrogradation  de  leur  solubilite  sous  l'influence  du  chlorure  de 

calcium,  Bui.  Soc.  chim.  [ser.  4]  13,  929-935. 
1913b  Lindet,  L.:  Sur  les  caseines  solubles  du  lait,  Ibid.  [ser.  4]  13,  1001-1006. 

1906  Lipschitz,  Arnold:  Ueber  den  Einfluss  der  Hautpflege  des  Milchviehs 

sowie  tiber  die  Einwirkung  einiger  Mineralstoffbeigaben  zum  Kraft- 

futter  auf  Milchergiebigkeit  und  Beschaffenheit  der  Milch,  Ber.  d. 

landwirtsch.  Inst.  Konigsberg  in  Pr.  7,  1-51;  through  Biedermann's 

Centralbl.  f.  Agriculturchem.  35  (1906),  545-549. 
1910a  Lipschiitz,  Alex.:  Untersuchungen  iiber  den  Phosphorhaushalt  des  wach- 

senden  Hundes,  Arch.  exp.  Path.  u.  Pharm.  62,  210-243. 
1910b  Lipschiitz,  Alex.:  Ueber  den  Phosphor  des  Kotes,  Ibid.  62,  244-252. 
1911a  Lipschiitz:  Alex.:  Zur  Physiologie  des  Phosphorhungers  im  Wachstum, 

Arch.  ges.  Physiol.  143,  91-98. 
1911b  Lipschiitz,  Alex.:  Die  biologische  Bedeutung  des  Kaseinphosphors  fur 

den  wachsenden  Organismus,  Ibid.  143,  99-108. 
(1911)  Lisbonne,  Marc:  (Wirkung  von  Chloriden  und  von  Phosphaten  auf  die 

Starkeverzuckerung  durch  Speichel  und  Pankreassaft),  Compt.  rend. 

Soc.  de  biol.  70,  207;  through  Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  41 

(1911),  730. 

1907  Lochhead,  A.  C,  and  W.  Cramer:  On  the  Phosphorus  Percentage  of 

Various  Samples  of  Protagon,  Biochemical  Jour.  2,  350-356. 
1897     Locke,  F.  S.:  Note  on  the  Influence  of  "Peptone"  on  the  Clotting  of  Milk 
by  Rennet,  Jour,  of  Exp.  Med.  2,  493-499. 

1910  Loeb,  Adam:  Ueber  den  Eiweissstoffwechsel  des  Hundes  und  iiber  die 

Abscheidung  der  Galle  bei  Fiitterung  mit  Eiweiss  und  Eiweissabbau- 
produkten,  mit  besonderer  Beriicksichtigung  der  zeitlichen  Verhaltnisse, 
Zeit.  Biol.  55  (N.  F.  37),  167-235. 

1911  Loeb,  Adam :  Ueber  den  Einfluss  der  Lecithinverabreichung  auf  Kalk  und 

Magnesiaausscheidung,  Intern.  Beitrage  Path.  u.  Ther.  der  Ernahrungs- 
storungen  3,  235-237. 
1911     Lob,  Walther:  Beitrage  zur  Frage  der  Glykolyse.     II.  Mitteilung.     Die 
Bedeutung  der  Phosphate  fiir  die  oxydative  Glykolyse,  Biochem.  Zeit. 
32,  43-58. 


PHOSPHORUS  METABOLISM  653 

1906  Lobisch,  Wilhelm:  Ueber  Nuclein-Eiweissverbindungen  unter  besonderer 
Berticksiehtigung  der  Nucleinsaure  der  Milchdriise  und  ihrer  ange- 
blichen  Beziehung  zur  Kase'inbildung,  Beitrage  z.  chem.  Physiol,  u. 
Path.  8,  191-209. 

1890  Lonnberg,  Ingolf :  Beitrage  zur  Kenntniss  der  Eiweisskorper  der  Nieren 

und  der  Harnblase,  Skand.  Arch.  Physiol.  3,  1-13. 
1904    Loevenhart,  A.  S.:  Ueber  die  Gerinnung  der  Milch,  Zeit.  physiol.  Chem. 

41,  177-205. 
1906-7  Loevenhart,  A.  S.,  and  C.  G.  Souder:  On  the  Effect  of  Bile  upon  the 

Hydrolysis  of  Esters  by  Pancreatic  Juice,  Jour.  Biol.  Chem.  2,  415-425. 

1891  Loew,  Oscar:  Ueber  die  physiologischen  Functionen  der  Phosphorsaure, 

Biol.  Centralbl.  11,  269-281. 

1899  Loew,  Oscar:  The  Physiological  Role  of  Mineral  Nutrients,  U.  S.  Dept. 

Agr.,  Div.  Vegetable  Physiol,  and  Path.,  Bui.  18.    60  pp. 

1910  Loewe,  Siegfried:  Ueber  den  Phosphorstoffwechsel  bei  Psychosen  und 

Neurosen,  Zeit.  ges.  Neurol,  u.  Psychiat.,  Originalien  4,  250-259. 

1911  Loewe,  Siegfried:  Ueber  den  Phosphorstoffwechsel  bei  Psychosen  und 

Neurosen.      2.  Mitteilung,  Ibid.  5,  445-456. 
1912a  Loewe,  Siegfried:  Zur  physikalischen  Chemie  der  Lipoide.       I.  Bezie- 

hungen  der  Lipoide  zu  den  Farbstoffen,  Biochem.  Zeit.  42,  150-189. 
1912b  Loewe,  Siegfried:  Zur  physikalischen  Chemie  der  Lipoide.  II.  Die  Bezie- 

hungen  der  Lipoide  zu  anderen  organischen  Substanzen  (Narkoticis, 

Hypnoticis,  u.  a.),  Ibid.  42,  190-204. 
1912c  Loewe,  Siegfried:  Zur  physikalischen  Chemie  der  Lipoide.  III.  Diffusion 

in  Lipoiden,  Ibid.  42,  205,  206. 
1912d  Loewe,  Siegfried:  Zur  physikalischen  Chemie  der  Lipoide.       IV.  Die 

Eigenschaften  von  Lipoidlosungen  in  organischem  Losungsmittel.  Ibid. 

42;  207-218. 

1904  Loewenheim,  Jul.:  Physiologisch  und  therapeutisch  Erfahrungen  mit 

der  organischen  Phosphor,  insbesondere  mit  Phytin,  Berlin,  klin.  Woch- 
enschr.  41,  1220-1223. 
(1911)  Lowenstein,  Joseph:  Zur  Behandlung  der  progressiven  Paralyse  mit 
Nukle'insaureinjektionen,  Ibid.  48,  714-716;  through    Jahresb.    u.     d. 
Fortschr.  d.  Thierchem.  41  (1911),  923. 

1900  Loewi,  Otto:  Beitrage  zur  Kenntniss  des  Nucleinstoffwechsels.      I.  Arch. 

exp.  Path.  u.  Pharm.  44,  1-23. 
1900-01  Loewi,  Otto:  Untersuchungen  fiber  den  Nucleinstoffwechsel.      II.  Mit- 
theilung,  Ibid.  45,  157-185. 

1909  London,  E.  S.:  Zum  Chemismus  der  Verdauung  in  tierischen  Korper. 

XXXVI.  Mitteilung.       Ueber  das  Verhalten  der  Nucleoproteide  im 
Magendarmkanal,  Zeit.  physiol.  Chem.  62,  451-454. 

1910  London,  E.  S.,  and  Alfred  Schittenhelm :  Verdauung  und  Resorption  von 

Nucleinsaure  im  Magendarmkanal.      I.  Mitteilung,  Ibid.  70,  10-18. 

1911  London,  E.  S.,  and  Alfred  Schittenhelm:  Verdauung  und  Resorption  von 

Nucleinsaure  im  Magendarmkanal.      II.  Mitteilung,  Ibid.  72,  459-462. 

1912  London,  E.  S.,  Alfred  Schittenhelm,  and  Karl  Wiener:  Verdauung  und 

Resorption  von  Nucleinsaure  im  Magendarmkanal.      III.  Mitteilung, 
Ibid.  77,  86-91. 

1905  Long,  J.  H.:  On  the  Specific  Rotation  of  Salts  of  Casein,  Jour.  Amer. 

Chem.  Soc.  27,  363-366. 
1906a  Long,  J.  H.:  Some  Investigations  on  Salts  of  Casein,  Ibid.  28,  372-384. 


654  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1906b  Long,  J.  H.:  On  the  Extraction  of  Fat  from  Feces  and  the  Occurrence 
of  Lecithin,  Ibid.  28,  704-706;  also  Science  N.  S.  24  (1906),  244,  245. 

1907a  Long,  J.  H.:  On  Some  Phenomena  Observed  in  the  Peptic  Digestion  of 
Caseins,  Jour.  Amer.  Chem.  Soc.  29,  223-230. 

1907b  Long,  J.  H. :  On  the  Combining  Power  of  Casein  with  Certain  Acids,  Ibid. 
29,  1334-1342. 

1908  Long,  J.  H.:  Observations  on  the  Stability  of  Lecithin,  Ibid.  30,  881-895. 
1908a  Long,  J.  H.,  and  Frank  Gephart:  On  the  Behavior  of  Emulsions  of  Leci- 
thin with  Metallic  Salts  and  Certain  Non-electrolytes,  Ibid.  30,  895-902. 

1908b  Long,  J.  H.,  and  Frank  Gephart:  On  the  Behavior  of  Lecithin  with  Bile 
Salts,  and  the  Occurrence  of  Lecithin  in  Bile,  Ibid.  30,  1312-1318. 

1906  Long,  J.  H.,  and  W.  A.  Johnson:  The  Phosphorus  Content  of  Feces  Fat, 

Ibid.  28,  1499-1503. 

1907  Long,  J.  H.,  and  W.  A.  Johnson:  Further  Observations  on  the  Nature  of 

Feces  Fat,  Ibid.  29,  1214-1220. 

1909  Lothrop,  Alfred  Peirce:  The  Effects  of  Bone  Ash  in  the  Diet  on  the 

Gastro-intestinal  Conditions  of  Dogs,  Amer.  Jour.  Physiol.  24,  297-324. 

1870  Lubavin,  N. :  Ueber  die  kiinstliche  Pepsin- Verdauung  des  Caseins  und  die 
Einwirkung  von  Wasser  auf  Eiweisssubstanzen,  Hoppe-Seyler's  Med. 
-Chem.  Unters.  463-485. 

1877  Lubavin,  N.:  Untersuchungen  iiber  der  chemischen  Natur  des  Nucleins, 
Ber.  deut.  chem.  Gesell.  10,  2237-2240. 

1889  Luciani,  Luigi:  Fisiologia  del  digiuno.  Studi  sull'  uomo.  (Das  Hun- 
gem.)       Florence.      157  pp.       German  edition  1890. 

1899  LUning,  Q.:  Die  anorganische  Bestandtheile  des  Pankreas,  Inaug.  Diss., 
Wtirzburg.       26  pp. 

1903  Liithje,  Hugo:  Ueber  die  Kastration  und  ihre  Folgen.      II.  Mitt.:  Einfluss 

der  Kastration  auf  den  Phosphorsaure-  und  Kalkstoffwechsel,  Arch, 
exp.  Path.  u.  Pharm.  50,  268-272. 

1904  Liithje,  H.,  and  CI.  Berger:  In  welcher  Form  kommt  aus  der  Nahrung 

retinierter   Stickstoff  im   Organismus   zur  Verwendung,  Deut.  Arch. 

klin.  Med.  81,  278-315. 
1901     Lumiere,  Auguste,  Louis  Lumiere,  and  F.  Perrin:  Sur  l'acide  glycero- 

phosphoreux  et  les  glycerophosphorites,  Compt.  rend.  Acad,  des  sci. 

139,  643,  644. 
1903     Lusena,  Gustavo:  Sul  contenuto  lecitinico  del  fegato,  dei  reni  e  del  cuore 

nella  degenerazione  adiposa  sperimentale,  Lo  Sperimentale  57,  29-46; 

through  Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  33   (1903),  90. 
1909     McCollum,  E.  V.:  Nuclein  Synthesis  in  the  Animal  Body,  Amer.  Jour. 

Physiol.  25,  120-141;  also  Wis.  Agr.  Exp.  Sta.  Research  Bui.  8  (1910). 
1913a  McCollum,  E.  V.,  and  Marguerite  Davis:  The  Influence  of  the  Compo- 
sition and  Amount  of  the  Mineral  Content  of  the  Ration  on  Growth,, 

Proc.  Soc.  Biol.  Chem.,  in  Jour.  Biol.  Chem.  14,  XL. 
1913b  McCollum,  E.  V.,  and  Marguerite    Davis:    The    Necessity    of    Certain 

Lipins  in  the  Diet  During  Growth,  Jour.  Biol.  Chem.  15,  167-175. 
1912     McCollum,  E.  V.,  and  S.  G.  Halpin:  Synthesis  of  Lecithins  in  the  Hen, 

Proc.  Soc.  Biol.  Chem.,  in  Jour.  Biol.  Chem.  11,  XIII,  XIV. 
1912     McCollum,  E.  V.,  J.  G.  Halpin,  and  A.  H.  Drescher:  Synthesis  of  Lecithin 

in  the  Hen  and  the  Character  of  the  Lecithins  Produced,  Jour.  Biol. 

Chem.  13,  219-224. 


PHOSPHORUS  METABOLISM  655 

1908  McCollum,  E.  V.,  and  E.  B.  Hart :  On  the  Occurrence  of  a  Phytin-splitting 

Enzyme  in  Animal  Tissues,  Ibid.  4,  497-500. 
1906a  McCrudden,  Francis  H.:  The  Composition  of    Bone    in    Osteomalacia, 

Amer.  Jour.  Physiol.  17,  32-34. 
1906b  McCrudden,  Francis  H.:  The  Effects  of  Castration  on  the  Metabolism  in 

Osteomalacia,  Ibid.  17,  211-217. 
1907     McCrudden,  Francis  H.:  The  Effect  of  Castration  on  Metabolism,  Proc. 

Soc.  Biol.  Chem.  1,  134,  135;  also  in  Jour.  Biol.  Chem.  4  (1908),  XL, 

XLI. 
1910a  McCrudden,  Francis  H.:  The  Effect  of  Castration  on  the  Metabolism, 

Jour.  Biol.  Chem.  7,  185-197. 
1910b  McCrudden,  Francis  H.:  Chemical  Analysis  of  Bone  from  a  Case  of 

Human  Adolescent  Osteomalacia,  Ibid.  7,  199-200. 

1912  McCrudden,  F.  H.,  and  Helen  L.  Fales:  Complete  Balance  Studies  of 

Nitrogen,  Sulphur,  Phosphorus,  Calcium  and  Magnesium  in  Intestinal 

Infantilism,  Jour,  of  Exp.  Med.  15,  450-456. 
1908a  MacLean,  Hugh:  Versuche  fiber  den  Cholingehalt  des  Herzmuskel-leci- 

thins,  Zeit.  physiol.  Chem.  57,  296-303. 
1908b  MacLean,  Hugh:  Ueber  das  Vorkommen  eines  Monaminodiphosphatids 

im  Eigelb,  Ibid.  57,  304. 
1909a  MacLean,  Hugh:  On  the  Nitrogen- Containing  Radicle  of  Lecithin  and 

Other  Phosphatides,  Biochemical  Jour.  4,  38-58;  240-257. 
1909b  MacLean,  Hugh :  On  the  Occurrence  of  a  Mon-amino-diphosphatide  Leci- 
thin-like Body  in  Egg  Yolk,  Ibid.  4,  168-174. 
1909c  MacLean,   Hugh:   Untersuchungen   liber   Eigelb-Lecithin,   Zeit.  physiol. 

Chem.  59,  223-229. 
1909d  MacLean,  Hugh:  Phosphatides  in  the  Light  of  Modern  Research,  British 

Med.  Jour.  1909,  II,  677,  678. 
1912a  MacLean,  Hugh:  The  Phosphatides  of  the  Kidney,  Biochemical  Jour.  6, 

333-354. 
1912b  MacLean,  Hugh:  On  the  Purification  of  Phosphatides,  Ibid.  6,  355-361. 
1912-13  MacLean,  Hugh:  On  Carnaubon,  Proc.  of  the  Physiol.  Soc,  in  Jour,  of 

Physiol.  45,  XVIII. 

1913  MacLean,  Hugh,  Die  Phosphatide  des  Herzens  und  anderer  Organe,  Bio- 

chem.  Zeit.  57,  132-142. 
1899     MacLeod,  J.  J.  R. :  Zur  Kenntniss  des  Phosphors  im  Muskel,  Zeit.  physiol. 

Chem.  28,  535-558. 
1906     MacLeod,  J.  J.  R.,  and  H.  D.  Haskins:  Some  Observations  on  the  Behav- 
ior of  the  Endogenous  Purin  Excretion  in  Man,  Jour.  Biol.  Chem.  2, 

231-242. 
1905a  Maestro,  Leone:  Fitina  e  Protilina.      Contribute  alio  studio  del  ricambio 

fosforganico,  La  Clinica  moderna  11,  517-522. 
1905b  Maestro,  Leone:  Sull'  assimilazione  della  fitina,  Lo   Sperimentale  59, 

456-458. 
1898     Magnani:  Le  iniezioni  endomuscolari  di  fosfoluteina  (lecitina  Serono)  in 

terapia  oculare,  Annali  di  ottalmologia  27,  569-571;  through  Stassano 

and  Billon,  1902d. 
1898    Magnus-Levy,  Adolf:  Ueber  die  Stoffwechsel  bei  akuter  und  chroniker 

Leukamie,  Arch.  path.  Anat.  u.  Physiol.  152,  107-130. 

1909  Magnus-Levy,  Adolf:  Der  Mineralstoffwechsel  in  der  klinischen  Patho- 

logie,  Zentralbl.  innere  Med.  30,  I,  503-505. 


656  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1869    Mai:  Ein  kleiner  Ffitterungsversuch  mit  phosphorsaurem  Kalk,  Jahresb. 

u.  d.  Fortschr.  d.  Agriculturchem.  11-12  (1868-9),  547. 
1894    Maier,  A.:      Ueber      seuchenartiges    Auftreten     der     Knochenweiche 

(Ehachitis)  bei  Schweinen  im  vorigen  Winter  und  Friihjahr  1894,  Ber- 
lin, tierarztlicher  Wochenschr.  543-546. 
1908a  Maillard,  L.  C:  Contribution  numerique  a  l'etude  de  l'excretion  urinaire 

de  l'azote  et  du  phosphore.       I.  Choix      des     sujets     et     techniques 

d'analyse,  Jour,  de  physiol.  et  de  path,  gener.  10,  985-1000. 
1908b  Maillard,  L.  C:  Contribution  numerique  a  l'etude  l'excretion  urinaire  de 

de  l'azote  et  du  phosphore.      II.  Resultats  d'ensemble,  Ibid.  10,  1017- 

1030. 
1909    Maillard,  L.  C:  Contribution  numerique  a.  l'etude  de  l'excretion  urinaire 

de  l'azote  et  du  phosphore.       III.  Discussion  des  resultats  moyens. 

IV.  Influence  de  l'exercice  musculaire.      V.  Recherche  de  l'influence 

eventuelle  du  vin,  Ibid.  11,  201-215. 
1902    Maillon,  CI.:  Contribution  a  l'etude  clinique    et    physiologique    de    la 

lecithine,  These,  Lyon. 

1902  Mainzer,  J.:  Experimented  Studien    iiber    die    Einwirkung    geistiger 

•Thatigkeit  auf  den  Harnstoffwechsel,  Monatsschr.  Psychiat.  u.  Neurol. 
11,  81-93. 

1903  Mainzer,  J.:  Stoffwechselstudien  iiber  den  Einfluss  gestiger  Tatigkeit 

und  protrahierten  Wachens,  Ibid.  14,  442-449. 
1884a  Mairet,  A.:  Recherches  sur  le  role  biologique  de  l'acide  phosphorique, 

Compt.  rend.  Acad,  des  sci.  99,  243-246. 
1884b  Mairet,  A.:  De  l'influence  du  travail  intellectuel  sur  l'elimination  de 

l'acide  phosphorique  par  les  urines,  Ibid.  99,  282-285. 
1884c  Mairet,  A. :  Recherches  sur  les  modifications  dans  la  nutrition  du  systeme 

nerveux  produites  par  la  manie,  la  lypemanie  et  l'epilepsie,  Ibid.  99, 

328-331. 
1884d  Mairet,  A.:  Recherches  sur  l'elimination  de  l'acide  phosphorique  chez 

l'homme  sain,  l'aliene,  l'epileptique,  et  l'hysterique,  Compt.  rend.  Soc. 

de  biol.  1884,  438-442;  461-465. 
1876    Makris,  C:  Studien  fiber  die  Eiweisskorper  der  Frauen-  und  Kuhmilch, 

Inaug.  Diss.,  Strassburg;  through  Jahresb.  fi.  d.  Fortschr.  d.  Thierchem. 

6,  (1876),  113-115. 
1902    Malcolm,  John:  Note  on  the  Percentage  Composition  of  Egg-Yolk,  Jour. 

of  Physiol.  27,  356-359. 

1904  Malcolm,   John:   On   the   Influence   of   Pituitary    Gland   Substance   on 

Metabolism,  Ibid.  30,  270-280. 

1900  Malengreau,  Fernand:  Deux  Nucleoalbumines  et  deux  Histones  dans  le 

Thymus,  La  Cellule,  17,  339-349. 

1901  Malengreau,  Fernand:  Sur  les  nucleins  du  thymus.  (Seconde  communi- 

cation.)      Ibid.  19,  283-309. 

1911  Malengreau,  F.,  and  Georges  Prigent:  Ueber  die  Geschwindigkeit  der 

Hydrolyse  der  Glycerinphosphorsaure,  Zeit.  physiol.  Chem.  73,  68-84. 

1912  Malengreau,  F.,  and  Georges  Prigent:  Ueber  Hydrolyse  und  Konstitution 

des  Lecithins,  Ibid.  77,  107-120. 

1905  Malerba,  P.:  Sul  ricambio  del  fosforo  nell'  encefalo,  Giornale  intemaz.  d. 

sci.  med.,  Napoli  27,  865-867. 

1906  Malerba,  P.:  Ueber  den  Stoffwechsel  des  Phosphors  im     Gehirn,    Zeit. 

angew.  Chem.  19,  917. 


PHOSPHORUS  METABOLISM  657 

1892  Malfatti,   Hans:   Beitrage  zur  Kenntniss   der  Nucleine,   Zeit.   physiol. 

Chem.  16,  68-86. 

1893  Malfatti,  Hans:  Bemerkung  zu  meinem  Aufsatze:  "Beitrage  zur  Kennt- 

niss der  Nucleine,"  Ibid.  17,  8,  9. 
1874    Mallet,  J.  W.:  Analysis  of  Buffalo  Bones,  Chem.  News  30,  211. 
1876    Maly,  M.:  Ueber  die  anderung  der  Reaction  (in  der  Losung  eines  Salz- 

gemisches)   durch  Diffusion  und  die  dar  durch  mogliche  Erklarung 

beim  Vorgange  der  Secretion  von  saurem  Harn  aus  alkalischem  Blute, 

Ber.  deut.  chem.  Gesell.  9,  164-172. 
1873    Maly,  Rich.,  and  Jul.  Donath:  Beitrage  zur  Chemie  der  Knochen,  Jour. 

prakt.  Chem.  115  (N.  F.  7),  413-441. 

1906  Manasse,  Armand :  Ueber  den  Gehalt  des  Eidotters  an  Lecithin,  Biochem. 

Zeit.  1,  246-252. 
1890    Manasse,  Paul:  Ueber  das  Lecithin  und  Cholesterin  der  rothen  Blut- 
korperchen,  Zeit.  physiol.  Chem.  14,  437-452. 

1895  Manasse,   Paul:   Ueber  zuckerabspaltende,   phosphorhaltige   Korper   in 

Leber  und  Nebenniere,  Ibid.  20,  478-488. 

1904  Mandel,  A.  R.,  and  G.  Lusk:  Stoffwechselbeobachtungen  an  einem  Falle 

von  Diabetes  mellitus,  mit  besonderer  Beriicksichtigung  der  Prognose, 
Deut.  Arch.  klin.  Med.  81,  472-492. 

1905  Mandel,  John  A.,  and  P.  A.  Levene:  Darstellung  und  Analyse  einiger 

Nucleinsauren.       XL   Mitt.   Ueber  die     Nucleinsaure   der  Kuhmilch- 

driise,  Zeit.  physiol.  Chem.  46,  155-158. 
1906a  Mandel,  John  A.,  and  P.  A.  Levene:  Darstellung  und  Analyse  einiger 

Nucleinsauren.      XII.  Mitt.  Ueber  die  Nucleinsaure  der  Niere,  Ibid. 

47,  140-142. 
1906b  Mandel,  John  A.,  and  P.  A.  Levene:  On  the  Pyrimidin  Bases  of  the 

Nucleic  Acid  Obtained  from  Fish  Eggs,  Jour.  Biol.  Chem.  1,  425,  426. 
1902     Mandel,  John  A.,  and  Horst  Oertel:     A  Further     Contribution    to     the 

Knowledge  of  the  Excretion  of  Organic  Phosphorus  in  the  Urine,  N. 

Y.  Univ.  Bui.  of  the  Med.  Sci.  I,  4,  165-170. 

1896  Marcuse,  Gotthelf :  Ueber  den  Nahrwerth  des  Caseins,  Arch.  ges.  Physiol. 

64,  223-248. 

1897  Marcuse,  Gotthelf:  Ueber  das  Verhalten  der  Phosphorausscheidung  bei 

Stoffwechselversuchen  mit  Casein,  Ibid.  67,  373-394. 
1905    Marfori,  Pio:  Sui  composti  organici  del  fosforo.   (Acido  fosfoglicerico 

e  glicerofosfati),  Archivio  di  fisiologia  2,  217-227;  through  Jahresb.   ii. 

d.  Fortschr.  d.  Thierchem.  35  (1905),  123. 
1908a  Marfori,  Pio:  Sui  composti  organici  del  fosforo.      (Acido  fosfoglicerico  e 

lecitina  per     iniezioni     sottocutanee.)       Ibid.  5,     207-216;     through 

Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  38  (1908),  588. 
1908b  Marfori,  Pio:  Sull'  assorbimento  e    sulla    assimilazione    dei    composti 

"organici"  del  fosforo,  Arch.  exp.  Path.  u.  Pharm.,  Suppl.,  Schmiede- 

berg  Festschrift,  378-388. 

1898  Marischler,  J.:  (Ueber  einen  Fall  von  iibermassiger  Schweissabsonder- 

ung  der  oberen  Korperhalfte  mit  einer  Untersuchung  des  Stoffwech- 
sels.)  Przeglad  lekarski  37,  No.  41  &  42;  through  Jahresb.  ii.  d. 
Fortschr.  d.  Thierchem.  29  (1899),  739,  740. 

1907  Mark,  H.:  Ueber  das  Jekorin,  Diss.,  Leipzig;  through  Jahresb.  ii.  d. 

Fortschr.  d  .Thierchem.  37  (1907),  465. 


658  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1895     Markotum,  K.:  Ueber  ein  phosphorhaltigen  Eiweisskorper  in  der  Schild- 

druse  im  Zusammenhang  mit  der  Frage  iiber  die  Function  derselben, 

St.   Petersburger   med.    Wochenschr.    1896,   Beilage   No.    1;   through 

Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  26  (1896),  518,  519. 
1911a  Marre,  Francis:  Contribution  a  1 'etude  de  l'utilisation  des  sels  mineraux 

du  lait  par  le  nourrisson,  Revue  d'hygiene  et  de  med.  infant.  10,  37-45. 
(1911b)  Marre,  Francis:  (Das  Lecithin  der  Butter  milch),  Revue  gener.  de  chim. 

pure  et  appl.  14,  56-58;  through  Jahresb.  ii.  d.  Fortschr.  d.  Thierchem. 

41  (1911),  187. 
1913     Marshall,  E.  K.,  Jr.:  On  the  Self-Digestion  of  the  Thymus,  Jour.  Biol. 

Chem.  15,  81-84. 
1911     Martin,  R.:  (Zusammensetzung  der  Schaf milch  des  Bezirks  Roquefort), 

Annal.  des  Falsific.  4,  86-88;  through  Zeit.  Unters.  d.  Nahrungs  und 

Genussmittel  25  (1913),  62,  63. 
1902a  Martinet,  Alfred:  Indications  et  contre-indications     de     la     medication 

phosphorique,  La  Presse  med.  Jan.   8  and  11,  1902;   through  Jour. 

Amer.  Med.  Assoc.  38  (1),  431. 
1902b  Martinet,  Alfred:  Les  nucleines  en  therapeutique,  Ibid.  10  (2),  1215-1218. 

1903  Martinet,  Alfred:  La  medication  phosphorique  dans  la  psychasthenic, 

Ibid.  11  (2),  805-807. 
1905     Martinet,   Alfred:   Pharmacologic   de   l'acide   phosphorique.     1:   Action 

digestive;  2.  Action  nerveuse  et  humerale,  Revue  gener.  des  sci.  16, 

568-575;  611-616. 
1910     Masing,  Ernst:  Ueber  das  Verhalten  der  Nucleinsaure  bei  der  Forchung 

des  Seeigeleis,  Zeit.  physiol.  Chem.  67,  161-173. 
1911a  Masing,   Ernst:   Chemische   Beitrage  zur  Blutregeneration,  Arch.   exp. 

Path.  u.  Pharm.  66,  71-83. 
1911b  Masing,  Ernst:  Ueber  eine  Beziehung    zwischen     Kernstoffgehalt    und 

Entwickelung,  Ibid.  75,  135-140. 
1913     Maslow,  M.  S.:  Ueber  die  biologische  Bedeutung  des  Phosphors  fur  den 

wachsenden  Organismus  und  seine  Einwirkung  auf  die  intrazellularen 

Fermente,  St.  Petersburger  med.  Wochenschr.  38,  97,  98. 

1904  Masoin,  P.:  Nouvelles  recherches  chimiques     sur     l'epilepsie,     Archives 

internat.  de  pharmacodynamic,  13,  387. 
1902     Massacin,  Cornel :  Ueber  den  Einfluss  des  lecithins  auf  den  Eiweissansatz, 

Deut.  med.  Wochenschr.  28,  756,  757. 
1913a  Masslow,  M.:  Ueber  die  biologische  Bedeutung  des  Phosphors  fur  den 

wachsenden  Organismus.       I.  Untersuchungen  iiber  den  Einfluss  des 

Phosphors  auf  die  Entwicklung  von  Tieren  und  auf  den  Phosphor-  und 

Stickstoffumsatz,  Biochem.  Zeit.  55,  45-62. 
1913b  Masslow,  M.:  Ueber  die  biologische  Bedeutung  des  Phosphors  fur  den 

wachsenden  Organismus.       II.  Untersuchung  der  Organe  auf  ihren 

Gehalt  an  Phosphor  und  intracellularen  Fermenten,  Ibid.  56,  174-194. 
1901     Massol  and  Gamel:  Du  role  des  hypophosphites  dans  le  medication  phos- 

phatee  et  hypophosphitee,  Bui.  Acad,  de  med.  de  Paris,  Oct.  8,  1901; 

through  Jour.  Amer.  Med.  Assoc.  37,  II,  1353. 
1910    Masuda,  Niro:  Ein  Beitrag  zur  Analyse  des  Gehirns,  insbesondere  iiber 

den  Cholesterin-  und  Fettsaurengehalt  desselben,  Biochem.  Zeit.  25, 

161-164. 
1897    Mathews,  Albert:  Zur  Chemie  der  Spermatozoen,  Zeit.  physiol.  Chem.  23, 

399-411. 


PHOSPHORUS  METABOLISM  659 

1910  Mathison,  G.  C:  The  Output  of  Organic  Phosphorus  in  Urine,  Biochem- 

ical Jour.  4,  274-279. 
1908    Matsushita,  T.:  Ueber  die  Aetiologie  der  Polyneuritis  gallinarum  und  der 

Beriberi,  Ber.  ii.  d.  14.  internat.  Kongr.  Hygiene  u.  Demographie  4, 

141-144. 
1901     Maurel,  E.:  Influence  des  variations  de  ralimentation  sur  les  quantites 

d'acide  phosphorique  et  de  chlorures  contenus  dans  Purine,  Compt. 

rend.  Soc.  de  biol.  53,  429-431. 

1904  Maurel,  E. :  E  valuation  approximative  de  la  quantite  minima  d'acide  phos- 

phorique urinaire  et  de  la  quantite  minima  de  cette  substance  neces- 

saire  a.  l'organisme    dans    les    conditions    de    la    ration    moyenne 

d'entretien,  Ibid.  56,  751-754. 
1891     Maxwell,  W.:  On  the  Behavior  of  the  Fatty  Bodies,  and  the  Role  of  the 

Lecithines,  during  Normal  Germination,  Amer.  Chem.  Jour.  13,  16-24. 
1893     Maxwell,  W.:  Movement  of  the  Element  Phosphorus  in  the  Mineral, 

Vegetable,  and  Animal  Kingdoms,  and  the  Biological  Function  of  the 

Lecithines,  Ibid.  15,  185-195. 
1913     Mayer,  Andre,  and  Georges  Schaeffer:    Recherches    sur    la    Constance 

lipocytique.       Teneur  des  tissus  en  lipoides  phosphores,  Compt.  rend. 

Acad,  des  sci.  157, 156-159. 
1907     Mayer,  Arthur:  Beitrage  zur  Kenntnis  des     Mineralstoffwechsels     der 

Phthisiker,  Deut.  Arch.  klin.  Med.  90,  408-424. 
1907    Mayer,  A.,  and  E.  F.  Terroine:   Sur  les  jecorines  naturelles  et  arti- 

ncielles,  Compt.  rend.  Soc.   de  biol.  62,  773-775. 
1896     Mayer,  Paul:  Ueber  den  Einfluss  von  Nuclein  und  Thyreoidinfutterung 

auf  die  Harnsaureausscheidung,  Deut.  med.  Wochenschr.  22,  186-188. 

1905  Mayer,  Paul:  Ueber  das  Verhalten  des  Lecithins  zu  den  Fermenten,  Ber- 

lin, klin.  Wochenschr.  42,  1102,  1103. 
1906a  Mayer,  Paul:  Ueber  die  Spaltung  der  lipoid  Substanzen  durch  Lipase  und 

liber  die  optischen  Antipoden  des  naturlichen  Lecithins,  Biochem.  Zeit. 

1,  39-52. 
1906b  Mayer,  Paul:  Ueber  Lecithinzucker  und  Jekorin  sowie  iiber  das  physikal- 

isch-chemische  Verhalten  des  Zuckers  im  Blut,  Ibid.  1,  81-107. 
1913     Mayesima,  Junichi:   Ueber   die   Resorption  der   Hefenucleinsaure  nach 

ausgedehnter  Resektion  des  Dunndarms  beim  Hunde,  Zeit.   physiol. 

Chem.  87,  418-422. 

1911  Medigreceanu,  Florentin:  Neuere  Arbeiten  iiber     die     Nuclease,     Med. 

Klinik.  Wien.,  7,  II,  1209,  1210. 

1911  Medigreceanu,  F.,  and  L.  Kristeller:  General  Metabolism  with  Special 
Reference  to  Mineral  Metabolism  in  a  Patient  with  Acromegaly  Com- 
plicated with  Glycosuria,  Jour.  Biol.  Chem.  9,  109-120. 

1910  Meier,  F.:  Ueber  den  Nucleinstoffwechsels  des  Schweines,  Inaug.  Diss., 
Giessen.       35  pp.       Through  Exp.  Sta.  Record  26   (1912),  363. 

1882  Meigs,  Arthur  V.:  Milk  Analysis,  Philadelphia  Med.  Times,  July,  1882, 

660-664. 

1883  Meigs,  Arthur  V.:  Proof  that  Human  Milk  Contains  only  About  One 

Percent  of  Casein;  with  Remarks  upon  Infant  Feeding,  Proc.  Philadel. 
Co.  Med.  Soc.  6,  92-108. 

1912  Meigs,  Edward  B.:  Contributions  to  the  General  Physiology  of  Smooth 

and  Striated  Muscle,  Jour.  Exp.  Zool.  13,  497-571. 


660  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1912    Meigs,  Edward  B.,  and  L.  A.  Ryan:  The  Chemical  Analysis  of  the  Ash 
of  Smooth  Muscle,  Jour.  Biol.  Chem.  11,  401-414. 

1905  Meinertz,  J.:  Zur  Kenntnis  des  Jecorins,  Zeit.  physiol.  Chem.  46,  376- 

382. 
1909    Meisen,  Jacob:  Experimentelle  Untersuchungen  uber  die  Wirkung  von 
Nucleinsaure  auf  Blut  und  Knochenmark,  Inaug.  Diss.,  Bonn.  28  pp. 

1911  Meisen,  Jacob :  Ueber  die  Wirkung  der  Nucleinsaure,  speziell  auf  Blut  und 

Knochenmark,  Med.  Klinik.  Wien.,  7,  II,  1946. 

1912  Mellanby,  J.:  The  Coagulation  of  Milk  by  Rennin,  Jour,  of  Physiol.;  45, 

345-362;  through  Chem.  Abs.  7  (1913),  1726,  1727. 
1872    Mendel,  E.:  Die  Phosphorsaure  im    Urin    von    Gehirnkranken,    Arch. 
Psychiat.  u.  Nervenkrankh.  3,  636-672. 

1906  Mendel,  Lafayette  B.:  The  Formation  of  Uric  Acid,  Jour.  Amer.  Med. 

Assoc.  46,  I,  843-846;  944-947. 
1907-8  Mendel,  Lafayette  B.,  and  Philip  H.    Mitchell:    Chemical    Studies    on 
Growth.       II.  The  Enzymes  Involved  in  Purine  Metabolism  in  the 
Embryo,  Amer.  Jour.  Physiol.  20,  97-116. 

1906  Mendel,   Lafayette  B.,  and  Frank  P.  Underhill:  Experiments   on  the 

Physiological  Action  and  Metabolism     of     Anhydro-oxy-methylene- 

diphosphoric  Acid  (Phytin  Acid),  Ibid.  17,  75-88. 
1903    Mendel,  Lafayette  B.,  Frank  P.  Underhill,    and    Benjamin    White:    A 

Physiological  Study  of  Nucleic  Acid,  Ibid.  8,  377-403. 
1909    Menten,  Maude  L.:  The  Distribution    of    Fat,    Chlorides,    Phosphates, 

Potassium  and  Iron  in  Striated  Muscle,  Trans.  Canadian  Inst.  8,  III, 

403-421. 

1911  Merck,  E.:  The  Glycerophosphates,  Annual  Report  25,  1-30.      About  68 

references. 

1912  Merck,  E.:  Lecithin,  Ibid.  26,  1-71.      About  525  titles  and  references. 

1907  Mesernitzy,  P.:  (Die  quantitative  Veranderung  des  Lecithins  im  Organis- 

mus  wahrend  der  Entwickelung.)       Russky  Wratsch,  1907,  302-304; 
through  Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  37  (1907),  513. 

1912  Messing,  Basia:  Ueber  einige  mineralische  Bestandteile  im  normalen 

und  pathologischen  Gehirn,     Inaug.     Diss.,    Zurich,    1912;     through 
Zentralbl.  Biochem.  u.  Biophys.  14  (1912),  133. 
1911     Mestrezat,  W.:    Composition    chimique    du    liquide    cephalo-rachidien 
normal.      Vraie  nature  de  cette  humeur,  Bui.  Soc.  chim.  9,  683-688. 

1913  Meyer,  Curt:  Zur  Kenntnis  des  Mineralstoffwechsels  bei  der  Rachitis, 

Jahrb.  Kinderheilk.      77,  28-52. 
1904a  Meyer,  Ludwig  F.:  Beitrage  zur  Kenntnis    des    Phosphorstoffwechsels, 

Zeit.  physiol.  Chem.  43,  1-10. 
1904b  Meyer,  Ludwig  F.:  Beitrage  zur  Kenntnis    des    Phosphorstoffwechsels, 

Beitrage  z.  wissensch.  Med.  u.  Chem.,  Fortschrift  zu  Ehren Ernst 

Salkowski,  261-263. 
1905    Meyer,  Ludwig  F.:  Zur  Kenntnis  des  idiopathischen  Oedems  des  Saug- 

lings,  Deut.  med.  Wochenschr.  31,  II,  1464-1467. 

1908  Meyer,  Ludwig  F.:  Zur  Kenntnis  des  Mineralstoffwechsels  im  Sauglings- 

alter,  Biochem.  Zeit.  12,  422-465. 

1909  Meyer,  Ludwig  F.:  Der  Mineralstoffwechsel  im  fruhen  Kindesalter,  Med. 

Klinik,  Berlin,  5,  I,  581-584. 
1898    Michaelis,  Leonor:  Beitrage  zur  Kenntniss  der    Milchsecretion,    Arch, 
mikroscop.  Anat.  51,  711-747. 


PHOSPHORUS  METABOLISM  661 

1913  Michaelis,  Leonor:  Die  allgemeine  Bedeutung  der  Wasserstoffionen- 
konzentration  fur  die  Biologie,  Oppenheimer's  Handbuch  der  Biochem. 
des  Mensch.  u.  der  Tiere,  Erganzungsband,  10-62. 

1908  Michaelis,  Leonor,  and  Peter  Rona:  Untersuchungen  liber  den  parenter- 

alen  Eiweissstoffwechsel.      I.  Arch.  ges.  Physiol.  121,  163-168. 

1896  Michel,  Charles:  Recherches  sur  la  nutrition  normale  du  nouveau-ne, 

L'Obstetrique  1,  140-158. 

1897  Michel,  Charles:  Sur  le  lait  de  femme  et  l'utilization  de  ses  materiaux 

nutritifs  dans  l'organisme  du  nouveau-ne  sain,  Ibid.  2.      518-533. 

1898  Michel,  Charles:  Composition  moyenne  du  lait  de  femme,  L'Union  phar- 

maceut.  39,  385-389. 

1899  Michel,  Charles:  Sur  la  composition  chimique  de  l'embryon  et  du  foetus 

humains  aux  differentes  periodes  de  la  grossesse,  Compt.  rend.  Soc.  de 
biol.  51,  422,  423. 

1900  Michel,  Charles:  Sur  la  composition  organique  et  minerale  du  foetus  et 

du  nouveau-ne,  L'Obstetrique  5.  252-260. 

1899  Michel,  Charles,  and  M.  Perret:  Etude  des  echanges  nutritifs  azotes  et 

mineraux  chez  un  nourrisson  de  2  mois  V2,  Bui.  Soc.  d'obstetrique  de 
Paris  2,  98-105. 
1906    Michel,  Charles,  and  M.  Perret:  La  ration  alimentaire  de  l'enfant  depuis 
sa  naissance  jusqu'  a  l'age  de  deux  ans,  Rev.  Soc.  d'hyg.  aliment,  et 
de  1'alimentation  rationnelle  de  l'homme  3,  209-362. 

1900  Micko,  K.:  Vergleichende  Untersuchungen  iiber  die    bei    Plasmon  und 

Fleischnahrung  ausgeschiedenen  Kothe,  Zeit.  Biol.  39,  430-450. 
1900    Micko,  K.,  P.  Mttller,  H.  Poda,  and  W.  Prausnitz:  Untersuchungen  iiber 

das  Verhalten  animalischer  Nahrungsmittel  im  menschlichen  Organ- 

ismus,  Ibid.  39,  277-278. 
1878    Miescher,  F.:  Die  Spermatozoen  einiger  Wirbeltiere.      Ein  Beitrag  zur 

Histochemie,  Verhandl.  d.  naturforsch.  Gesell.  z.  Basel  6,  138-208. 
1881    Miescher,  F.:  Ueber  das  Leben  des  Rheinlaches  im  Siisswasser,  Arch. 

Anat.  u.  Physiol./ anat.  Abt.,  193-218. 

1896  Miescher,  F.:  Physiologisch-chemische  Untersuchungen  iiber  die  Lachs- 

milch,  (Edited  by  O.  Schmiedeberg)  Arch.  exp.  Path.  u.  Pharm.  37, 
100-155. 

1897  Miescher,  F.:  Die  histo-chemischen  und  physiologischen  Arbeiten  von 

Friedrich  Miescher  gesammelt  und  herausgegeben  von  seinen  Freund- 

en.      Leipzig,  2  Vols.  138  and  543  pp. 
1904    Migliaccio,  Alessandro:  La  lecitina  in  terapia  infantile.     Osservazioni 

sull'  azione  del  rimedio  in  alcune  malattie  dei  bambini,  La  Pediatria 

12,  753-769. 
1911    Mihara,  Shinji:  Beitrage  zur    Kenntnis    der    Fermente  der  Stierhoden, 

Zeit.  physiol.  Chem.  75,  443-455. 

1898  Milesi,  C:  Di  un  corpo  fosforato  isolato  dell'  albume  dell'  novo  pres- 

entante  i  caratteri  chimici  d'un  mucoide,  Bollett.  della  Soc.  med.  chir. 
di  Pavia,  1898,  No.  3,  p.  184;  through  Jahresb.  ii.  d.  Fortschr.  d.  Thier- 
chem.  28  (1898),  38,  39. 
1911     Milkowicz,   G.   S.:    (Experimental   Increase   of  Phagocytosis),   Russky 
Wratsch,  1911,  No.  22,  943-946;  through  Chem.  Abs.  6  (1912),  252. 

1909  Miller,  J.  R.,  and  Walter  Jones:  Ueber  die  Fermente  des  Nucleinstoff- 

wechsels  bei  der  Gicht,  Zeit.  physiol.  Chm.  61,  395-404. 


662  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1896  Milroy,  T.  H.:  Ueber  die  Eiweiss-Verbindungen  der  Nucleinsaure  und 
Thyminsaure  und  ihre  Beziehung  zu  den  Nuclei'nen  und  Paranucleinen, 
Zeit.  physiol.  Chem.  22,  307-326. 

1908  Milroy,  T.  H. :  Changes  in  the  Chemical  Composition  of  the  Herring  Dur- 

ing the  Reproductive  Period,  Biochemical  Jour,  3,  366-390. 

1898  Milroy,  T.  H.,  and  J.  Malcolm:  The  Metabolism  of  the  Nucleins  under 

Physiological  and  Pathological  Conditions.  Part  I.  Jour,  of  Physiol. 
23,  217-239. 

1899  Milroy,  T.  H.,  and  J.  Malcolm:  The  Metabolism  of  the  Nucleins.       Part 

II.  Further  Observations  on  Excretion  in  Leucocythaemia.  Intra- 
cellular Metabolism  in  the  Granular  Leucocytes,  Ibid.  25,  105-130. 

1912  Minami,  D.:  Ueber  den  Einfluss  des  Lecithins  und  der  Lipoide  auf  die 
Diastase  (Amylase),  Biochem.  Zeit.  39,  355-380. 

1898  Minkowski,  O.:  Untersuchungen  zur  Physiologie  und  Pathologie  der 
Harnsaure  bei  Saugethieren,  Arch.  exp.  Path.  u.  Pharm.  41,  375-420. 

1878  Miroczkowski :  Ueber  den  Phosphorsauregehalt  im  Schafs-,  Kalbs-  und 
Hunde-Serum,  Centralbl.  med.  Wissensch.  16,  353,  354. 

1911  Mitra,  Mariano:  (Presence  of  Nuclease  and  Connectivase  in  the  Gastric 
Contents  of  Infants),  Folia  clinica  3,  274-278;  through  Chem.  Abs.  5 
(1911),  3591  and  7  (1913),  640. 

1902  Mitulesco,  J. :  Beitrage  zum  Studium  des  Stoffwechsels  in  der  chronischen 

Tuberkulose,  Berlin,  klin.  Wochenschr.  1027-1029;  1054-1056;  1073- 
1075;   1096-1100. 

1903a  Mitulesco,  J.:  Die  Vortheile  der  combinierten  Behandlung  bei  der  chron- 
ischen Tuberkulose.  Vortrag.  Deut.  med.  Wochenschr.  29,  367-369; 
385,  386. 

1903b  Mitulesco,  J.:  Beitrage  zum  Studium  des  Stoffwechsels  in  der  Tuber- 
kulose, Zeit.  Tuberkulose  4,  515-537. 

1909  Mochi,  Alberto:  L'eliminazione  dell'  N,  P,  Ca,  Mg,  nei  conigli  digiuni 

trattati  con  iniezioni  di  estratto  di  ipofisi,  Atti  r.  Accad.  d.  fisiocrit. 
Siena  218  (1  of  ser.  5), -835-856. 

1910  Mochi,  Alberto:  II  ricambio  dell'  N,  P,  e  Ca  nei    conigli     trattati    con 

iniezioni  di  estratto  di  ipofisi,  Rivista  d.  patol.  nerv.  e  ment.  15,  457- 

476. 
1901     Mochizuki,  Junichi:  Ueber  die  Resorption  von  Eiweisskorpern  von  der 

Schleimhaut  des  Dickdarmes  nach  Versuchen  mit  Thymus-klystieren, 

Arch.  Verdauungskrankh.  7,  221-233. 
1901     Modica,  O.,  and  E.  Audenino:   (Einfluss  des  Zentralnervensystems  auf 

den  organischen   Stoffwechsel),  Archivio  di  psichiat.,   sci.   penali   ed 

antropol.  22,  Fasc.  4-5;  through  Jahresb.  ii.  d.  Fortschr.  d.  Thierchem. 

32  (1902),  672,  673. 
1869     Mors  and  Muck:  Beitrage  zur  Kenntniss  der  Osteomalazie,  Deut.  Arch. 

klin.  Med.  5,  485-504. 

1903  Mohr,  L. :  Ueber  das  Ausscheidungsvermogen  der  kranken  Niere,  Zeit. 

klin.  Med.  51,  331-348. 
1905     Moll,   Leopold:  Beitrag  zur  Ernahrungstherapie  der  mit  Phosphaturie 

(Calcarurie)   einhergehenden  Neurosen  im  Kindesalter,  Prager  med. 

Wochenschr.  30,  582-585. 
1909     Moll,   Leopold:  Die  klinische  Bedeutung  der  Phosphorausscheidung  im 

Harn  beim  Brustkind,  Jahrb.  Kinderheilk.  69  (19  of  ser.  3),  129-152; 

304-332;   450-478. 


PHOSPHORUS  METABOLISM  663 

1912     Molyakov,   M.   G.:    (Administration   of    Sodium   Nucleinate   in    Scarlet 

Fever),  Russki  Wratch  .11    (1912),  301-303;   through  Chem.  Abs.   7, 

(1913),  1921. 
(1911)  Monrad:  Kaseinklumpen  im  Kinderstuhl  im  Zusammenhang  mit  Roh- 

milchernahrung,  Monatsschr.    f.    Kinderheilk.    10,    244-246;    through 

Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  41  (1911),  298. 
1910     Monvoisin,   A.:    (Die   Zusammensetzung  krankhaft   veranderter   Milch, 

insebesondere  der  Milch  tuberkuloser  Kuhe),  Jour,  de  Pharm.  et  de 

Chim.  [7]  2,  23-30;  through  Zeit.  Unters.  d.  Nahrungs-  u.  Genussmittel 

23  (1912),  25. 
1901     Moore,  Benjamin,  and  William  H.  Parker:  On  the  Functions  of  the  Bile 

as  a  Solvent,  Proc.  Roy.  Soc.  of  London  68,  64-76. 
1908     Moore,  Benjamin,  Herbert  E.  Roef,  Robert  E.  Knowles:  The  Effects  of 

Variations  in  the  Inorganic  Salts  and  the  Reactivity  of  the  External 

Medium  upon  the  Nutrition,  Growth  and  Cell  Division  in  Plants  and 

Animals,  Biochemical  Jour.  3,  279-312. 
1895a  von  Moraczewski,  Waclaw:  Verdauungsproducte  des    Caseins    und     ihr 

Phosphorgehalt,  Zeit.  physiol.  Chem.  20,  28-51. 
1895b  von  Moraczewski,  Waclaw:  Ueber  den  Chlor-  und  Phosphorgehalt  des 

Blutes  bei  Krebskranken,  Arch.  path.  Anat.  u.  Physiol.  139,  385-405. 
1896     von  Moraczewski,  Waclaw:  Ueber  den  Chlor-    und    Phosphorgehalt    des 

Blutes  bei  krankhaften  Zustanden,  Ibid.  146,  424-452. 
1897a  von  Moraczewski,  Waclaw:  Die  Mineralbestandtheile  der  menschlichen 

Organe,  Zeit.  physiol.  Chem.  23,  483-496. 
1897b  von  Moraczewski:  Stoffwechselversuch  bei  Diabetes  mellitus,  CentralbL 

innere  Med.  18,  921-932. 
1897c  von  Moraczewski,  Waclaw:  Stoffwechseluntersuchungen     bei     Carcinom 

und  Chlorose,  Zeit.  klin.  Med.  33,  385-431. 
1898a  von  Moraczewski,  Waclaw:   Stoffwechselversuch  bei  Diabetes  mellitus, 

Ibid.  34,  59-88. 
1898b  von  Moraczewski,  Waclaw:     Stoffwechselversuche     bei     Leukamie     und 

Pseudoleukamie,  Arch.  path.  Anat.  u.  Physiol.  151,  22-52. 
1899     von  Moraczewski,  Waclaw:  Ueber  die  Ausscheidung  der  Harnbestand- 

theile  bei  Fieberbewegungen,  Ibid.  155,  11-43. 
1900a  von  Moraczewski,  Waclaw:     Stoffwechsel    bei    Lungenentziindung    und 

Einfiuss  der  Salze  auf  denselben,  Zeit.  klin.  Med.  39,  44-92. 
1900b  von  Moraczewski,  Waclaw:  Stoffwechselversuche  bei  schweren  Anamien, 

Arch.  path.  Anat.  u.  Physiol.  159,  221-252. 

1901  von  Moraczewski,  Waclaw:  Stoffwechsel  bei  Akromegalie  unter  der  Be- 

handlung  mit  Sauerstoff,  Phosphor,  etc.,  Zeit.  klin.  Med.  43,  336-360j 

611-616. 
(1902)  von  Moraczewski,  Waclaw:  (Ueber  den  Verlauf  des  Fiebers  beim  Men- 

schen  mit  ausgeschnittener  Milz),  Gazeta  lekarska  22,  868;  through 

Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  32  (1902),  746. 
1903-4  von  Moraczewski,  Waclaw :  Ueber  Ausscheidung  von  Oxalsaure,  Indican 

und  Aceton  bei  Diabetes  unter  dem  Einfiuss  der  Nahrung,  Zeit.  klin. 

Med.  51,  475-501. 
1905     von  Moraczewski,  Waclaw:  Ein  Beitrag  zur  Kenntnis  der  Phosphaturie, 

CentralbL  innere  Med.  26,  401-414. 

1902  Moreigne,  H.:  Action  du  jus  de  raisin  sur  l'organisme,  Archives  de  med. 

exper.  et  d'anat.  14,  343-377. 


664  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1909  Morel,  L.:  Les  parathyroides  dans  l'osteogenese.  (Premiere  note.)  Compt. 

rend.  Soc.  de  biol.  67,  780-782. 

1910  Morel,   L.:   Les   parathyroides   dans   l'osteogenese.        (Deuxieme  note.) 

Ibid.  68,  163-165. 
1906     Morgan,  A.,  C.  Beger,  and  G.  Fingerling:  Weiterer  Untersuchungen  iiber 

die  Wirkung  der  einzelnen  Nahrstoffe  auf  die  Milchproduction,  Land- 

wirtsch.  Versuch.  Stat.  64,  93-242. 
1901     Morichau-Beauchant,   P.   E.   R,:  Etude   therapeutique   sur  la   lecithine, 

These  de  Paris.       141  pp. 
1885     Mosetig-Moorhof :  Milchsaure  als  Zerstorungsmittel  pathogener  Gewebe, 

Centralbl.  Chirurgie,  193-196. 
1853     Mosler,  Friedrich:  Beitrage  zur  Kenntniss  der  Urinabsonderung  bei  ge- 

sunden,  schwangeren  und  kranken  Personen,  insbesondere  quantitative 

Bestimmung  der  phosphorsauren  Verbindungen.  Inaug.  Diss.,  Giessen. 
1899     Mosse,  Prosper  and  Oulie:  Influence  de  l'ovariotomie  double  et  de  l'in- 

gestion  d'ovaires  sur  quelques  elements  de  la  secretion  urinaire  chez 

la  chienne,  Compt.  rend.  Soc.  de  biol.  51,  447-449. 

1911  Moszkowski,  Max:  Meine  Erfahrungen  iiber  Prophylaxe  der  Beriberi  in 

Hollandisch-Neuguinea,  Arch.  Schiffs-  u.  Tropenhygiene  15,  653-661. 
1899    Mott,  F.  W.,  and  J.  O.  Wakelin  Barratt:  Observations  on  the  Chemistry 
of  Nerve  Degeneration,  Archives  of  Neurol.,  1899,  I,  346-358. 

1904  Mott,  F.  W.,  Arthur  Edmunds,  and  W.  D.  Halliburton:  Regeneration  of 

Nerves,  Proc.  Roy.  Soc,  March  1904. 

1901  Mott,  F.  W.,  and  W.  D.  Halliburton:  The  Chemistry  of  Nerve-degenera- 

tion, Phil.  Trans,  of  the  Roy.  Soc.  of  London  194  B  (1901),  437-466; 
Jour,  of  Physiol.  26,  XXV,  XXVI;  through  Jahresb.  u.  d.  Fortschr.  d. 
Thierchem.  31  (1901),  558. 

1902  Mott,  F.  W.,  and  W.  D.  Halliburton:  Regeneration  of  Nerves,  Ann.  Rept. 

of  the  British  Assoc.  1902. 
1902a  Mouneyrat,  A.:  De  l'arsenic  et  du  phosphore  organiques  dans  le  traite- 

ment  de  la  tuberculose  pulmonaire.       (Extrait.)       Compt.  rend.  Acad. 

des  sci.  134,  667,  668. 
1902b  Mouneyrat,  A.:  Sur  une  nouvelle  medication  arsenio-phosphoree   (His- 

togenol)  dans  le  traitement  de  la  tuberculose  pulmonaire,  Compt.  rend. 

Soc.  de  biol.  54  (4  of  ser  11),  314,  315. 

1905  Moussu,  G.,  and  J.  A.  W.  Dollar:  Diseases  of  Cattle,  Sheep,  Goats  and 

Swine,  New  York,  1905.      785  pp. 
1902     Miiller,  Erich:  Beitrag  zum  Kalkstoffwechsel  des  Sauglings,  Verhandl.  d. 

Gesell.  Kinderheilk.  19-20  (1902-3),  221-226. 
1911     Miiller,  Erich:  TJeber  Ernahrung  debiler  Kinder  mit  molkenreduzierter 

Milch  an  der  Hand  von  Stoffwechseluntersuchungen,  Jahrb.  Kinder- 
heilk.     Supplement,  73,  252-262. 
1913     Miiller,  Erich,  and  Ernst  Schloss:  Beitrage  zur  Kenntnis  des  Stoffwech- 

sels  besonders  der  Mineralien  im  Sauglingsalter.      I.  Einleitung,  Ibid. 

77,  635-639. 
1884     Miiller,  Friedrich:  Ueber  den  normal  en  Koth  des  Fleischfressers,  Zeit. 

Biol.  20,  327-377. 
1889    Miiller,  Friedrich:  Stoffwechseluntersuchungen  bei  Krebskranken,  Zeit. 

klin.  Med.  16,  496-549. 
1873    Miiller,  Jakob  Worm:  Zur  Kenntniss  der  Nucleine,  Arch.  ges.  Physiol.  8, 

190-194. 


PHOSPHORUS  METABOLISM  665 

1913  Miiller,  Johannes,  and  H.  Reinbach:  Ueber  Maskierung  des  Blutfettes 
und  der  Blutlipoide  sowie  uber  Verdauungslipamie  beim  Menschen, 
Zeit.  physiol.  Chem.  86,  469-483. 

1897  Miiller,  Martin :  Ueber  den  Gehalt  der  menschlichen  Muskeln  an  Nucleon, 

Zeit.  physiol.  Chem.  22,  561-566. 
1900     Miiller,  Paul:  Ueber  den  organischen  Phosphor  der  Frauenmilch-  und  der 

Kuhmilchfazes,  Zeit.  Biol.  39,  451-481. 
1908     Miiller,  W.,  and  G.  von  Wendt:  Wie  futtert  der  Landwirt  zweckmassig 

Rubenblatter  ?       Berlin.   .  31  pp. 

1898  Muggia,  Alberto:  Sulle  iniezioni  di  lecitina  e  di  tuorlo  d'uovo  nella  cura 

dell'  anemia  ed  atrepsia  infantile,  La  Pediatria  6,  302-308. 

1911  Mulier,  Rechla:  Ueber  Phosphorsaureausscheidung  bei  Lungentuberku- 

lose,  Inaug.  Diss.,  Zurich.  40  pp. 

1908  Munk,  Fritz:  Ueber  "lipoid  Degeneration,"  Arch.     path.     u.     Anat.     u. 

Physiol.  194  (4  of  ser.  19),  527-565. 
1887     Munk,  Immanuel:  Ueber  die  Ausfuhr  des   Stickstoffs  und  der  Asche- 
bestandteile  durch  den  Harn,  Berlin,  klin.  Wochenschr.  430-433. 

1894  Munk,  Immanuel:  Beitrage  zur     Stoffwechsel-     und     Ernahrungslehre, 

Arch.  ges.  Physiol.  58,  309-408. 

1895  Munk,  Immanuel :  Ueber  den  Einfluss  angestrengter  Korperarbeit  auf  die 

Ausscheidung  der  Mineralstoffe  und  der  Aetherschwefelsauren,  Arch. 

Anat.  u.  Physiol.,  physiol.  Abt.,  385-387. 
1904     Nakayama,  M.:  Ueber  das  Erepsin,  Zeit.  physiol.  Chem.  41,  348-362. 
1842    Nasse,  H.:  Ueber  die  Bestandtheile  der  Knochen  in  einigen  Krankheiten, 

Jour.  f.  prakt.  Chem.  (Erdmann)  27,  274-280. 

1899  Nemser,  M.  M.  H.:  Sur  la  question  de  savoir  comment  les  nucleines  se 

comportent  dans  l'inanition,  Archives  des  sci.  biol.  7,  221-232. 
1908a  Nerking,  Joseph:  Beitrage  zur  Kenntniss  des  Knochenmarks,  Biochem. 

Zeit.  10,  167-191. 
1908b  Nerking,  Joseph:  Die  Verteilung  des  Lecithins  im  tierischen  Organismus, 

Ibid.  10,  193-203, 

1909  Nerking,  Joseph:  Narkose  und  Lezithin,  Munch,  med.  Wochenschr.  56,. 

II,  1475-1478. 

1908  Nerking,  J.,  and  E.  Haensel:  Der  Lecithingehalt  der  Milch,  Biochem. 
Zeit.  13,  348-353. 

1899a  Nesbitt,  Beattie:  On  the  Presence  of  Cholin  and  Neurin  in  the  Intes- 
tinal Canal  during  its  Complete  Obstruction,  Proc.  Amer.  Physiol.  Soc* 
in  Amer.  Jour.  Physiol.  2,  VIII. 

1899b  Nesbitt,  Beattie:  A  Research  on  Autointoxication,  Jour,  of  Exp.  Med.  4, 
1-18. 

1866-71  Nessler,  J.:  Knochenbruchigkeit  beim  Rindvieh,  Verhandl.  d.  natur- 
wissensch.  Vereins,  Karlsruhe,  3-5,  23,  24. 

1873  Nessler,  J.:  Untersuchungen  der  Knochen  von  Knochenbruchigen  Rind- 
vieh, Landwirtsch.  Versuch.  Stat.  16,  187-192. 

1900  Netter,  L.:  E  changes  nutritifs   dans  l'allaitement  artificiel,  These   de 

Paris.      79  pp. 

1912  Neubauer,  H.,  and  G.  Hillkowitz:  Der  Kalk-  und  Phosphor sauregehalt 

und  Nahrwert  des  im  Jahre  1911  geenteten  Heues  und  Strohs,  Land- 
wirtsch. Zeit.  Rheinprovinz  13,  111,  112. 
1908    Neuberg,  Carl :  Zur  Frage  der  Konstitution  des  "Phytins,"  Biochem.  Zeit. 
9,  557-560. 


666  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1909     Neuberg,  Carl:  Notiz  iiber  Phytin,  Ibid.  16,  406-410. 

(1911)  Neuberg,  Carl:  Versuche  mit  Jodocitin,  Ther.  d.  Gegenwart  52,  359,  360; 

through  Jahresb.  u.  d.  Fortschr.  d.  Thierchem.  41  (1911),  890. 
1907     Neuberg,  C,  and  B.  Brahn:  Ueber  die  Inosinsaure,  Biochem.  Zeit.   5, 

438-450. 

1911  Neuberg,  C,  and  E.  Kretschmer:  Weiteres  iiber  kiinstliche  Darstellung 

von     Kohlenhydratphosphorsaureestern     und     Glycerinphosphorsaure, 

Ibid.  36,  5-14. 
1910a  Neuberg,  C,  and  H.  Pollak:  Ueber  Kohlenhydratphosphorsaureester.  I. 

Ueber  Saccharosephosphorsaure,  Ibid.  23,  515-517. 
1910b  Neuberg,   C,   and  H.   Pollak:   Ueber  Kohlenhydrat-Phosphorsaureester. 

II.  Ueber  Saccharoseschwefelsaure  und     die     Phosphorylierung     von 

Eiweiss,  Ber.  deut.  chem.  Gesell.  43,  II,  2060-2068. 
1910c  Neuberg,  C,  and  H.  Pollak:  Ueber  Phosphorsaure-  und  Schwefelsaure- 

ester  von  Kohlenhydraten,  Biochem.  Zeit.  26,  514-528. 
1910d  Neuberg,  C,  and  H.  Pollak:  Ueber  die  Phosphorylierung  von  Eiweiss  und 

die  Bindung  des  Phosphors  in  den  naturlichen  Phosphorproteinen,  Ibid. 

26,  529-534. 

1898  Neumann,  Albert:  Zur  Kenntniss  der  Nucleinsubstanzen,  Arch.  Anat.  u. 

Physiol.,  physiol.  Abt.,  374-378. 

1899  Neumann,  Albert :  Verf ahren  zur  Darstellung  der  Nucleinsauren  a  und  ft 

und  der  Nucleothyminsaure,  Ibid.  1899,  Suppl.  552-555. 

1893a  Neumann,  J.:  Ueber  die  Einwirkung  des  dem  Futter  beigegebenen  phos- 
phorsauren  Kalkes  auf  den  Aschengehalt  der  Milch,  Milch-Zeitung  22, 
701-704. 

1893b  Neumann,  J.:  Tierphysiologische  Untersuchungen.  I.  Beitrag  zur 
Kenntniss  des  Assimilation  anorganischen  Nahrstoffe  im  Tierkorper, 
Jour.  f.  Landwirtsch.  41,  343-380. 

1894  Neumann,  J.:  Tierphysiologische  Untersuchungen.  II.  Ueber  den  Einfluss 
des  phosphorsauren  und  kohlensauren  Kalkes  auf  die  Korperge- 
wichtszunahme,  Ibid.  42,  33-67. 

1906  Neumann,  Rudolf  Otto:  Untersuchungen  iiber  die  Einwirkung  des 
Protylins  auf  die  Phosphorausscheidung  des  Menschen,  Munch,  med. 
Wochenschr.  53,  1558-1561. 

1894a  Neumann,  Siegfried:  Quantitative  Bestimmung  des  Calciums,  Magne- 
siums und  der  Phosphorsaure  im  Harn  und  Koth  bei  Osteomalacic, 
Arch.  Gynakol.  47,  202-223. 

1894b  Neumann,  Siegfried:  Ueber  die  Verhaltnisse  der  Ausscheidung  von  Cal- 
cium, Magnesium  und  Phosphorsaure  bei  Osteomalacie,  Ungar.  Arch. 
Med.  3,  276-298. 

1896  Neumann,  Siegfried:  Weitere  Untersuchungen  iiber  die  Stoffwechselver- 
haltnisse  des  Calciums,  Magnesiums,  der  Phosphorsaure  und  des  Nitro- 
gens bei  puerperaler  Osteomalacie,  mit  besonderer  Rucksicht  auf  die 
durch  die  Kastration  und  andere  therapeutische  Eingriffe  verursachten 
Veranderungen  des  Stoffwechsels,  Arch.  Gynakol.  51,  130-185. 

1902  Neumann,  Siegfried,  and  Bernbard  Vas:  Ueber  den  Einfluss  der  Ovar- 
iumpraparate  auf  den  Stoffwechsel,  Monatsschr.  Geburtshilfe  u. 
Gynakol.  15,  433-451. 

1912  Newman,  L.  F.,  G.  W.  Robinson,  E.  T.  Hainan,  and  H.  A.  D.  Neville: 

Some  Experiments  on  the  Relative  Digestibility  of  White  and  Whole 
Meal  Breads,  Jour,  of  Hygiene  12,  119-143. 


PHOSPHORUS  METABOLISM  667 

1908     Nichols,  J.  B.:  Acid  Intoxication,  Washington  Med.  Ann.  7,  133-149. 
1886     Nissen,  Franz:  Ueber  das  Verhalten  der  Kerne     in     der     Milchdruse 
wahrend  der  Absonderung,  Arch,  mikroskop.  Anat.  26,  337-342. 

1912  Nizzi,  F.:    Le  bilan  du  phosphore,    de    la    lecithine    et    des    graisses, 

Encephale  7,  II,  245. 
1911     Njegovan,  Vladimir:  Beitrage  zur  Kenntniss     der     pflanzlichen     Phos- 
phatide, Zeit.  physiol.  Chem.  76,  1-26. 

1913  Njegovan,  Vladimir:  Enthalt  die  Milch  Phosphatide?     Biochem.  Zeit.  54, 

78-82. 
1898     Nolf,  P.:  Des  nucleines,  Annales  de  l'lnst.  Pasteur  12,  361-368. 

1898  Noll,  Alfred:  Ueber  die   Bildung  von  Lavulinsaure   aus   Nucleinsaure, 

Zeit.  physiol.  Chem.  25,  430-433. 

1899  Noll,  Alfred:  Ueber  die  quantitativen  Beziehungen  des  Protagons  zum 

Nervenmark,  Ibid.  27,  370-397. 

1907  von  Noorden,  Carl:  Metabolism  and  Practical  Medicine.       Vol.  I.  The 

Physiology  of  Metabolism.  Vols.  II  and  III.  The  Pathology  of 
Metabolism.       Chicago.       452,  525,  and  1320  pp. 

1883-4  North,  W.:  The  Influence  of  Bodily  Labor  upon  the  Discharge  of  Nitro- 
gen, (Abstract)  Proc.  Roy.  Soc.  of  London  36,  11-17. 

1904  Novi,  Ivo:  L'eliminazione  dei  fosfati  durante  la  cura  antirabica  e  la  sua 
modificazione  per  opera  della  terapia  fosfoglicerica,  Memoire  d.  r» 
Accad.  d.  sci.  d.  Institute  di  Bologna  [6]  1,  157-177. 

1908  Novi,  Ivo:  Terapia  della  fosfaturia  con  preparati  organici  del  fosforo- 

glicerofosfato  sodico  e  fitina),  Ibid.  [6]  5,  97-108. 

1909  Novi,  Ivo:  Fitina  e  glicerofosfati.       Nota  II.  Azione  sul  lavoro  mus- 

colare,  Ibid.  [6]  6,  301-316. 
1874     Odenius,  M.  V.,  and  J.     Lang:     Fall     af    lymphorrhoea,     pachydermia 

lymphorrhagica  af  Dr.  M.  V.  Odenius,  i  Lund,  Nordiskt  Medicinskt 

Arkiv  6,  No.  13;   through  Jahresb.  ii.   d.  Fortschr.   d.   Thierchem.   5 

(1875),  128,  129. 
1899     Odenius,  R. :  Nagra  undersokningar  ofver  en  nukleoproteid  ur  mjolk- 

korteln,  Upsala  Lakaref.  Forh.  (N.  F.)  5,  582-588;  through  Jahresb. 

ii.  d.  Fortschr.  d.  Thierchem.  30  (1900),  39. 
1899     Oechsner,  De  Coninck:  Sur  l'elimination  de  l'azote  et  du  phosphore  chez 

les  nourrissons,  Compt.  rend.  Acad,  des  sci.  129,  223,  224. 
1901     von  Oefele,     Freih.:     Trippelphosphate     im     Stuhlgang,     Pharmazeut. 

Zentralhalle  42,  191-198. 
1907     von  Oefele,  Freih.:  Bioson  als  Phosphornahrmittel  auf  Grund  von  Kot- 

untersuchungen,  Deut.  med.  Presse  11,  27,  28. 
1909     Oeri,  Felix:  Ein  Beitrag  zur  Kenntnis  des  Phosphorsaure-  und  Kalk- 

stoffwechsels  beim  erwachsenen  gesunden  Menschen,  Zeit.  klin.  Med. 

67,  288-306;  307-318. 
1898-9  Oertel,  Horst :  Beitrag  zur  Kenntniss  der  Ausscheidung  des  organisch 

gebundenen  Phosphors  im  Harn,  Zeit.  physiol.  Chem.  26,  123-170. 
1911-12  Ogata,  Masakiyo:  Ueber  das  Wesen  der  Rachitis  und  Osteomalacic, 

Beitrage  z.  Geburtshilfe  u.  Gynakol.  17,  23-54. 
1912     Onodera,  Nakamura  and  Tateno :  Untersuchung  iiber  den  Stoffwechsel  bei 

den   Kakkekranken  je   nach  der   Verabreichung   von   poliertem   Reis 

Oder  nicht  poliertem  Reis,  Mitt.  Med.  Ges.  Tokio  26,  No.  23,  4,  5. 


668  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1904     Orgler,  Arnold:  Ueber  das  Vorkommen  eines  protagonartigen  Korpers 

in  der  Nebennieren,  Beitrage  z.  wissensch.  Med.  u.  Chem.,  (Festschrift 

f.  Ernst  Salkowski),  285-288. 
1902     Osborne,  Thomas:  Bestimmung  des  Schwefels  in  den  Proteinkorpern, 

Zeit.  anal.  Chem.  41,  25-35. 
1900a  Osborne,  Thomas  B.,  and  George  F.  Campbell:  The  Nucleic  Acid  of  the 

Embryo  of  Wheat  and  its  Protein  Compounds,  Jour.  Amer.  Chem.  Soc. 

22,  379-413;  also  23rd  Ann.  Rept.  Conn.  Agr.  Exp.  Sta.,  New  Haven  (for 

1899),  305-339. 
1900b  Osborne,  Thomas  B.,  and  George  F.  Campbell:  The  Proteids  of  the  Egg 

Yolk,  Jour.  Amer.  Chem.  Soc.  22,  413-422;  also  23rd  Ann.  Rept.  Conn. 

Agr.  Exp.  Sta.,  New  Haven,  (for  1899),  339-348. 
1900c  Osborne,  Thomas  B.,  and  George  F.  Campbell:  The  Protein  Constituents 

of  Egg  White,  Jour.  Amer.  Chem.  Soc.  22,  422-450. 

1911  Osborne,  Thomas  B.,  and  H.  H.  Guest:  Hydrolysis  of  Casein,  Jour.  Biol. 

Chem.  9,  333-353. 
1902a  Osborne,  Thomas  B.,  and  Isaac  F.  Harris:  The    Nucleic    Acid    of    the 

Wheat  Embryo,  Ann.  Rept.  Conn.  Agr.  Exp.  Sta.  for  1901,  365-430. 
1902b  Osborne,  Thomas  B.,  and  Isaac  F.  Harris:  Die  Nucleinsaure  des  Weizen- 

embryos,  Zeit.  physiol.  Chem.  36,  85-133. 

1908  Osborne,  Thomas  B.,  and  F.  W.  Heyl:  The  Pyrimidine  Derivatives  of 

Nucleic  Acid,  Amer.  Jour.  Physiol.  21,  157-161. 

1909  Osborne,  Thomas  B.,  and  D.  Breese  Jones:  Hydrolysis  of  Vitellin  from 

the  Hen's  Egg,  Ibid.  24,  153-160. 

1911a  Osborne,  T.  B.,  and  L.  B.  Mendel:  Feeding  Experiments  with  Isolated 
Food-Substances,  Carnegie  Inst.,  Pub.  156,  parts  1  &  2. 

1911b  Osborne,  T.  B.,  and  L.  B.  Mendel:  The  Role  of  Different  Proteins  in  Nu- 
trition and  Growth,  Science  34,  722-732. 

1912a  Osborne,  T.  B.,  and  L.  B.  Mendel:  Feeding  Experiments  with  Fat-Free 
Food  Mixtures,  Jour.  Biol.  Chem.  12,  81-89. 

1912b  Osborne,  T.  B.,  and  L.  B.  Mendel:  Ueber  Futterungsversuche  mit  isolier- 
ten  Nahrungssubstanzen,  Zeit.  physiol.  Chem.  80,  307-382. 

1912c  Osborne,  T.  B.,  and  L.  B.  Mendel:  The  Role  of  Gliadin  in  Nutrition,  Jour. 
Biol.  Chem.  12,  473-510. 

1912d  Osborne,  T.  B.,  and  L.  B.  Mendel:  Maintenance  Experiments  with  Iso- 
lated Proteins,  Ibid  13,  233-276. 

1913  Osborne,  T.  B.,  and  L.  B.  Mendel:  The  Relation  of  Growth  to  the  Chemi- 
cal Constituents  of  the  Diet,  Ibid.  15,  311-326. 

1901-2  Osborne,  W.  A.:  Caseinogen  and  its  Salts,  Jour,  of  Physiol.  27,  398-406. 

1908  Ostertag,  and  Zuntz:  Untersuchungen  iiber  die  Milchsecretion  des 
Schweines  und  die  Ernahrung  der  Ferkel,  Landwirtsch.  Jahrbiicher 
37,  201-260. 

1899  Oswald,  Ad.:  Die  Eiweisskorper  der  Schilddriise,  Zeit.  physiol.  Chem.  27, 
14-49. 

1902  Oswald,  Ad. :  Die  Chemie  und  Physiologie  des  Kropf  es,  Arch.  path.  Anat. 
u.  Physiol.  169,  444-479. 

1912  Otolski,  and  Biernacki:  Ueber  die  Phosphatide  in  den  Organen  der  mit 

getoteten  Tuberkelbacillen  geimpften  Kaninchen,  Biochem.  Zeit.   41, 
375-385. 
1906     Otolsky,  S.  W.:  Das  Lecithin  des  Knochenmarks,  Ibid.  4,  124-153. 


PHOSPHORUS  METABOLISM  669 

1886    Ott,  Adolf:  Ueber  einige  die  Phosphate  des  Harnes  betreffende  Ver- 

haltnisse,  Zeit.  physiol.  Chem.  10,  1-10. 
1903    Ott,  Adolf:  Zur  Kenntniss  des   Stoffwechsels  der  Mineralbestandteile 

beim  Phthisiker,  Zeit.  klin.  Med.  50,  432-440. 
1862    Pagenstecher:  Ein  Beitrag  zur  Statistik  des  Kaiserschnittes  nebst  einem 

Anhang:  Ueber  Osteomalacie,  Monatsschr.  Geburtzkunde  19,  111. 
1894    Pages,  C:  Physiologie  de  la  matiere  minerale  du  lait.      These  de  Paris, 

1894;  through  Milch-Zeitung  25,  86,  87  and  the  following  review. 
1888    Paijkull,  Lincoln:  Ueber  die  Schleimsubstanz  der  Galle,  Zeit.  physiol. 

Chem.  12, 196-210. 
1913    Paladino,  Raffaele:  Untersuchungen  iiber    einige    Veranderungen    des 

Stoffwechsels  bei  Tieren  nach  Exstirpation  der  Schilddruse  und  der 

Parathyroiden,  Biochem.  Zeit.  50,  497-507. 
{1900)  Panek,  K.:  (Ueber  das  Verhaltniss  des  Phosphorsauregehaltes  des  Harns 

zum  Kalk-  und  Magnesiagehalt  bei  der  sogenannten  Phosphaturie), 

Przeglqd  lekarski  39,  1;  through  Jahresb.  u.  d.  Fortschr.  d.  Thierchem. 

30  (1900),  772,  773. 
1902a  Panella,  Amilcare:  L'acido  fosfocarnico  dei  muscoli  dopo  la  morte,  Arch- 

ivio  di  farmacol.  e  terapeut.  10,  323-361. 
1902b  Panella,  Amilcare:     L'acido     fosfocarnico     nella     sostanza     cerebrale, 

Giornale  d.  reale  Accad.  di  med.  di  Torino  65,  398-400. 
1902c  Panella,  Amilcare:  L'acido  fosfocarnico  del  sangue,  Archivio  di  farmacol. 

e  terapeut.  10,  439. 
1903a  Panella,  Amilcare:  L'acide  phosphocarnique  dans  la  substance  cerebrale, 

Archives  ital.  de  biol.  39,  260-262. 
1903b  Panella,  Amilcare:  L'acide  phosphocarnique  des  muscles  apres  la  mort. 

Ibid.  39,  263-283. 
1903c  Panella,  Amilcare:  L'acide  phosphocarnique  du  sang,  Ibid.  39,  283-291. 
1903d  Panella,  Amilcare:  L'acide  phosphocarnique  dans  le  testicule,  Ibid.  39, 

441,  442. 
1903e  Panella,  Amilcare:  L'acide  phosphocarnique  des  muscles  blancs  et  des 

muscles  rouge,  Ibid.  39,  443-451. 
1903f   Panella,   Amilcare:   L'acido   fosfocarnico   dei   muscoli   bianchi   e   rossi, 

Archivio  di  farmacol.  e  terapeut.  11,  91-103. 
1903g  Panella,  Amilcare:  L'acide  phosphocarnique  dans  la  substance  nerveuse 

centrale,  Archives  ital.  de  biol.  39,  452-470. 
1903h  Panella,  Amilcare:  L'acido  fosfocarnico  nella  sostanza  nervosa  centrale, 

Giornale  d.  reale  Accad.  di  med.  di  Torino,  66,  423-443. 
1903i    Panella,  Amilcare:  Sul  contenuto  di  acqua  e  di  nucleone  dei  testicoli  di 

cavallo,  Archivio  di  farmacol.  e  terapeut.  11,  291-302. 
1904a  Panella,  Amilcare:  Sur  le  contenu  d'eau  et  de  nucleone  des  testicules  de 

cheval,  Archives  ital.  de  biol.  42,  289-297. 
1904b  Panella,  Amilcare:  Eau  et  nucleone  des  muscles  lisses,  Ibid.  42,  368-382. 
1904c  Panella,  Amilcare:  L'acqua  ed  il  nucleone  della  milza,  Archivio  di  fis- 

iologia  1.      539-549. 
1906a  Panella,  Amilcare:  II  nucleone  e  l'acqua  del  cervello  in  animali  a  digiuno, 

Archivio  di  farmacol.  speriment.  5,  70-76. 
1906b  Panella,  Amilcare:  Sul  rapporto  fra  quantita  di  sostanza  usata  e  quanti- 
ty di  nucleone  determinata,  Ibid.  5,  216-219. 


670  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1902  Panzer,  Theodor:  Ueber  das  Verhalten  von  unterphosphorigsauren  Cal- 
cium im  thierischen  Korper,  Zeit.  Untersuch.  d.  Nahrungs-  u.  Genuss- 
mitt.  5,  11-14. 

1906  Panzer,  Theodor:  Ueber  das  sogenannte  Protagon  der  Niere,  Zeit. 
physiol.  Chem.  48,  519-527. 

1870  Papillon,  Fernand:  Recherches  experimentales  sur  les  modifications  de  la 
composition  immediate  des  os,  Compt.  rend.  Acad,  des  sci.  71,  372-374. 

1870-1  Papillon,  Fernand:  Recherches  experimentales  sur  les  modifications  de 
la  composition  immediate  des  os,  Jour,  de  l'anat.  et  de  la  physiol.  7* 
152-163. 

1873  Papillon,  Fernand:  Recherches  experimentales  sur  les  modifications  de  la 
composition  immediate  des  os,  Compt.  rend.  Acad,  des  sci.  76,  352-355. 

1890  Pappel,  A.,  and  H.  Droop  Richmond:  The  Milk  of  the  Gamoose,  Jour,  of 
the  Chem.  Soc,  Trans.,  57,  754-760. 

1877  Paquelin  and  Joly:  Des  pyrophosphates  en  therapeutique;     leur     mode 

d'action,  Compt.  rend.  Acad,  des  sci.  85,  410-412. 

1878  Paquelin  and  Joly:  Du  role  physiologique  des  hypophosphites,  Ibid.  86, 

1505,  1506. 
1913     Parnas,  J.:  Ueber  die  gesattigte  Fettsaure  des  Kephalins,.Biochem.  Zeit. 
56,  17-20. 

1908  Parrozzani,  A.:  Influenza  di  quantito  progressive  di  concimi  fosfatici  sul 

contenuto  in  sostanze  organische  fosforate  ed  azotate,  e  sul  rapporto 
fra  fosforo  ed  azote  dei  semi  mais,  Staz.  sperim.  agrar.  ital.  41,  729- 
738. 

1909  Parrozzani,   A.:    (The  Relationship   of  Organic  Phosphorus   to  Amido- 

nitrogen  and  to  Non-proteid  Nitrogenous  Compounds  in  Ripe  Seeds), 

Rend.  Soc.  chim.  Ital.  2nd  ser.,  1,  205-207;  through  Exp.  Sta.  Record  25 

(1911),  432. 
1894     Pasqualis,  G.:  SulP  assorbimento  e  l'eliminazione  dell'  acido  fosfoglicer- 

ico  e  suo  ricerca  nelle  orine  e  nel  sangue,  Annali  di  chim.  e  farmacol., 

20  Agosto,  1894;  through  Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  24 

(1894),  283-285. 
1905     Passon,  Max:  Zur  Beifutterung  mit  phosphorsaurem  Kalk,  Jour.  f.  Land- 

wirtsch.  53,  113-134. 
1913     Patein,  G.:  Histones  et  nucleohistones;  leur  recherche  dans  les  liquides 

de  l'organisme,  Jour,  de  pharm.  et  de  chim.  7,  55-60. 
1895-6  Paton,  D.  Noel :    On  the  Relationship  of  the  Liver    to    Fats,    Jour,    of 

Physiol.  19,  167-216. 

1898  Paton,  D.  Noel:  The  Phosphorus  Compounds  of  the  Muscle  and  Genitalia 

of  the  Salmon,  and  their  Exchanges,  Fishery  Bd.  of  Scotland,  Rept.  of 

Investigations  on  the  Life  Hist,  of  Salmon,  143-155. 
1897-8  Paton,  D.  Noel,  with  Francis  D.  Boyd,  James  C.  Dunlop,  A.  Lockhart 

Gillespie,  G.  Lovell  Gulland,  E.  D.  W.  Greig,  S.  C.  Mahalanobis,  and 

M.  I.  Newbigin:  The  Physiology  of  the  Salmon  in  Fresh  Water,  Jour. 

of  Physiol.  22,  333-356. 
1900     Paton,  D.  Noel,  J.  C.  Dunlop,  and  R.  S.  Aitchison:  Contributions  to  the 

Study  of  the  Metabolism  of  Phosphorus  in  the  Animal  Body,  Ibid.  25, 

121-224. 

1899  Paton,  D.  Noel,  J.  C.  Dunlop,  and  Ivison  Macadam:  On  the  Modifications 

of  the  Metabolism  Produced  by  the  Administration  of  Diphtheria 
Toxine,  Ibid.  24,  331-335. 


PHOSPHORUS  METABOLISM  671 

1910  Patta,  Aldo:  Osservazioni  intorno  al  comportamento  degli  ipofosfiti  nell' 
organismo,  Archivio  di  farmacol.  speriment.  9,  1-7;  through  Jahresb. 
ii.  d.  Fortschr.  d.  Thierchem.  40  (1910),  1385. 

1912  Patta,  Aldo;  Nuove  ricerche  intorno  alia  influenza  della  lecitina  sul 
bilancio  azotato  e  fosforato,  Ibid.  13,  515-528. 

1904  Patten,  A.  J.,  and  E.  B.  Hart:  The  Nature  of  the  Principal  Phosphorus 

Compound  in  Wheat  Bran,  Amer.  Chem.  Jour.  31,  564-572;  also  N.  Y. 

Agr.  Sta.  Bui.  No.  250. 
1876a  Pavy,  F.  W.:  The  Effect  of  Prolonged  Muscular  Exercise  on  the  System, 

Lancet,  1876,  I,  319,  320;  353-356;  392-394;  429-431;  466-468. 
1876b  Pavy,  F.  W.  The  Effect  of  Prolonged  Muscular  Exercise  upon  the  Urine 

in  Relation  to  the  Source  of  Muscular  Power,  Ibid.  1876,  II,  815-818; 

887-889. 
1877     Pavy,  F.  W.:  The  Effect  of  Prolonged  Muscular  Exercise  upon  the  Urine 

in  Relation  to  the  Source  of  Muscular  Power,  Ibid.  1877,  I,  42-44. 

1895  Pekelharing,  C.  A.:  Ueber  die  Beziehung  des  Fibrinferments  aus  dem 

Blutserum  zum  Nucleoproteid,  welches   aus  dem   Blutplasma  zu  er- 
halten  ist,  Centralbl.  Physiol.  9,  102-111. 

1896  Pekelharing,  C.  A.:  Ueber  das  Vorhandensein  eines  Nucleoproteids  in 

Muskeln,  Zeit.  physiol.  Chem.  22,  245-247. 
1902     Pekelharing,  C.  A.:  Mittheilungen  iiber  Pepsin,  Ibid.  35,  8-30. 
1914     Pekelharing,  C.  A.:  Ueber  den  Einfluss  von  Phosphatiden  auf  die  Blut- 

gerinnung,  Ibid.  89,  22-38. 

1905  Pende,  N.:  II  sangue  nell'  osteomalacia,  Bullett.  d.  reale  Accad.  med.  di 

Roma,  31,  161-198. 
1882     Penzoldt,  F.,  and  R.  Fleischer:  Experimentelle  Beitrage  zur  Pathologie 

des  Stoffwechsels  mit  besonderer  Beriicksichtigung  des  Einflusses  von 

Respirationsstorungen,  Arch.  path.  Anat.  u.  Physiol.  87,  210-262. 
1902     Percival,  A.  L.:  Sur  les  variations  du  phosphore  mineral,  conjugue  et 

organique,  dans  tissus  animaux,  Compt.  rend.  Acad,  des  sci.  135,  1005- 

*007. 
1908     Peritz,  Georg:  Lues,  Tabes  und  Paralyse  in  ihren  atiologischen  und  ther- 

apeutischen  Beziehungen  zum  Lecithin,  Berlin,  klin.  Wochenschr.  45, 

I,  53-56. 
1908-9a  Peritz,  Georg:  Ueber  die  Beziehung  der  Lues,  Tabes  und  Paralyse  zum 

Lecithin,  Deut.  Zeit.  Nervenheilk.  36,  87-94. 
1908-9b  Peritz,  Georg:  Ueber  das  Verhaltniss  von  Lues,  Tabes  und  Paralyse 

zum  Lecithin,  Zeit.  exp.  Path.  u.  Ther.  5,  607-621. 
1910     Peritz,  Georg:  Zur  Pathologie  der  Lipoide,  Ibid.  8,  255-278. 

1892  Petit,  P.:  Distribution  et  etat  du  fer  dans  l'orge,  Compt.  rend.  Acad,  des 

sci.  115,  246-248. 

1893  Petit,  P.:  Sur  une  nucleine  vegetale,  Ibid.  116,  995-997. 

1873  Petrowsky,  D.:  Zusammensetzung  der  grauen  und  der  weissen  Substanz 
des  Gehirns,  Arch.  ges.  Physiol.  7,  367-370. 

1906  Petry,  Eugen:  Ueber  die  Einwirkung     des     Labferments     auf    Kasein, 

Beitrage  z.  chem.  Physiol,  u.  Path.  8,  339-364. 
1866     von  Pettenkofer,  Max,  and  Carl  Voit:  Untersuchungen  iiber  den  Stoff- 

verbrauch  des  normalen  Menschen,  Zeit.  Biol.  2,  459-573. 
1889     Peyer,   Alexander:   Die   Phosphaturie,    Sammlung   klinischer   Vortrage, 

Volkmann,  Innere  Med.  No.  112  (whole  No.  336). 


672  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1904  Pfaundler,  Meinhard:  Ueber  die  Elemente  der  Gewebsverkalkung  und 
ihre  Beziehung  zur  Rachitisfrage,  Jahrb.  Kinderheilk.  60,  123-177. 

1899  Pfeiffer,  Theodor,  and  Wilhelm  Scholz:  Ueber  den  Stoffwechsel  bei  Par- 
alysis agitans  und  in  Senium  uberhaupt  (mit  Beriicksichtigung  des 
Einflusses  von  Schilddriisen-tabletten),  Deut.  Arch.  klin.  Med.  63,  368- 
422. 

1874  Piccard,  J.:  Ueber  Protamin,  Guanin  und  Sarkin  als  Bestandtheile  des 
Lachssperma,  Ber.  deut.  chem.  Gesell.  7,  1714-1719. 

1910  Pighini,  Giacomo:  Ueber  die  Bestimmung  der  enzymatischen  Wirkung 

der  Nuclease  mittels  "optischer  Methode,"  Zeit.  physiol.   Chem.  70, 
85-93. 

1911  Pighini,  Giacomo:  Ueber  die  Esterase  und  Nuclease  des  Serums  bei  ver- 

schiedenen  Formen  von  Geisteskrankheiten,  Biochem.  Zeit.  33,  190-217. 

1912  Pighini,  G.,  and  Flaminio  Nizzi:   Chemische  und  biochemische  Unter- 

suchungen  liber  das  Nervensystem  unter  normalen  und  pathologischen 

Bedingungen.      III.  Mitt.  Aufsuchung  der  Esterase  und  der  Lecithase 

in  der  normalen  und  pathologischen  Cerebrospinal  fliissigkeit,  Ibid. 

42,  145-149. 
1906    Pignatti,  Augusto:  Influenza  dei  composti  fosforati  organici  sulla  ferrat- 

ina  e  sul  suo  contenuto  in  fosforo,  Bollett.  delle  sci.  med.  di  Bologna  77 

(6  of  ser.  8),  230-243;  through  Jahresb.  u.  d.  Fortschr.  d.  Thierchem.  36 

(1906),  585. 
1898     Pinzani:  (Experimentelle  Untersuchungen  liber  den  Einfluss  der  Kastra- 

tion  auf  den  Stoffwechsel  und  die  Blutbeschaffenheit),  Archiv.  ost.  e 

gin.,  1898;  through  Centralbl.  Gynakol.  23,  1311. 
1897    Piatt,  Charles:  The  Normal  Urine,  Jour.  Amer.  Chem.  Soc.  19,  382-384. 
1903    Plenge,  H.:  Ueber  die  a-nucleinsaures  Natron  losende  Wirkung  einiger 

Mikroorganismen,  Zeit.  physiol.  Chem.  39,  190-198. 

1908  Plimmer,  R.  H.  Aders:  The  Proteins  of  Egg-yolk,  Jour,  of  the  Chem. 

Soc.  93,  1500-1506. 

1913a  Plimmer,  R.  H.  Aders:  The  Metabolism  of  Organic  Phosphorus  Com- 
pounds. Their  Hydrolysis  by  the  Action  of  Enzymes,  Biochemical 
Jour.  7,  43-71. 

1913b  Plimmer,  R.  H.  Aders:  The  Hydrolysis  of  Organic  Phosphorus  Com- 
pounds by  Dilute  Acid  and  by  Dilute  Alkali,  Ibid.  7,  72-80. 

1906  Plimmer,  R.  H.  A.,  and  W.  M.  Bayliss:  The  Separation  of  Phosphorus 
from  Caseinogen  by  the  Action  of  Enzymes  and  Alkali,  Jour,  of 
Physiol.  33,  439-461. 

1909-10  Plimmer,  R.  H.  A.,  Maxwell  Dick,  and  Charles  C.  Lieb:  A  Metabolism 
Experiment  with  Special  Reference  to  the  Origin  of  Uric  Acid,  Ibid. 
39,  98-117. 

1909  Plimmer,  R.  H.  A.,  and  R.  Kaya:  The  Distribution  of  Phosphoproteins  in 

Tissues.      Part  II,  Ibid.  39,  45-51. 
1913    Plimmer,  R.  H.  A.,  and  Harold  J.  Page:  An  Investigation  of  Phytin,  Bio- 
chemical Jour.  7,  157-174. 

1908  Plimmer,  R.  H.  Aders,  and  F.  H.  Scott:  A  Reaction  Distinguishing  Phos- 

phoprotein  from  Nucleoprotein  and  the  Distribution  of  Phosphopro- 
teins in  Tissues,  Jour,  of  the  Chem.  Soc,  Trans.,  93,  1699-1721. 

1909  Plimmer,  R.  H.  Aders,  and  F.  H.  Scott:  The  Transformations  in  the 

Phosphorus  Compounds  in  the  Hen's  Egg  during  Development,  Jour, 
of  Physiol.  38,  247-253. 


PHOSPHORUS  METABOLISM  673 

1871    Plosz,  P.:  Ueber  das  chemische  Verhalten  der  Kerne  der  Vogel-  und 

Schlangenblutkorperchen,  Hoppe-Seyler's  Med.  chem.  Unters.,  461-462. 
1873    Plosz,  P.:  Ueber  die  eiweissartigen  Substanzen  der  Leberzelle,  Arch.  ges. 

Physiol.  7,  371-390. 
1900    Poda,  H.,  and  W.  Prausnitz:  Ueber  Plasmon,  ein  neues  Eiweisspraparat, 

Zeit.  Biol.  39,  279-312. 
1889     Pohl,  Julius:  Bemerkungen  fiber  kfinstlich  dargestellte  Eiweissnucle'ine, 

Zeit.  physiol.  Chem.  13,  292-297. 

1884  Politis,  Georgios:  Ueber  das  Verhaltnis  der  P205  zum  N  im  Harn  bei 

Fiitterung  mit  Gehirnsubstanz,  Zeit.  Biol.  20,  193-214. 

1906  Pollak,  Emil:  Die  Nucleoproteide  in  der  Behandlung  septischer  Krank- 

heitsformen,  Arch.  Gynakol.  89,  479-521. 

1885  Pommer,  Gustav:  Untersuchungen  iiber  Osteomalacie  und  Rachitis  nebst 

Beitragen  zur  Kenntniss  der  Knochenresorption  und  -Apposition  in 
verschiedenen  Altersperioden  und  der  durchbohrenden  Gefasse,  Leip- 
zig.      506  pp. 
1894    Popoff,  P.  M.:  Ueber  die  Einwirkung  von  eiweissverdauenden  Fermenten 
auf  die  Nucleinstoffe,  Zeit.  physiol.  Chem.  18,  533-539. 

1907  Porges,  O.,  and  E.  Neubauer:  Physikalisch-chemische  Untersuchungen 

iiber  das  Lecithin  und  Cholesterin,  Biochem.  Zeit.  7,  152-177. 
1905    Posner,  Edward  R.,  and  William  J.  Gies:  Protagon  a  Mechanical  Mixture 

of  Substances  or  a  Definite  Chemical  Compound?      Jour.  Biol.  Chem. 

1,  59-112. 
1900     Posternak,  S.:  Contribution  a.  l'etude  chimique  de  l'assimilation  chloro- 

phylliene.      Sur  le  premier  produit  d'organisation  de  l'acide  phosphor- 

ique  dans  les  plantes  a,  chlorophylle,  avec  quelques  remarques  sur  la 

role  physiologique  de  l'inosite,  Revue  gener.  de  botanique    12,    5-24; 

64-73. 
1903a  Posternak,   S.:   Sur  un  nouveau  principe  phospho-organique  d'origine 

vegetale,  la  phytine,  Compt.  rend.  Soc.  de  biol.  55,  1190-1192. 
1903b  Posternak,  S.:  Sur  la  matiere  phospho-organique  de  reserve  des  plantes 

a  chlorophylle,  Compt.  rend.  Acad,  des  sci.  137,  202-204. 
1903c  Posternak,  S.:  Sur  les  proprietes  et  la  matiere    phospho-organique    de 

reserve  des  plantes  a  chlorophylle,  Ibid.  137,  337-339. 
1903d  Posternak,  S.:  Sur  la  constitution    de    l'acide    phospho-organique     de 

reserve  des  plantes  vertes  et  sur  le  premier  produit  de  reduction  du 

gaz  carbonique  dans  l'acte  de  l'assimilation  carbonique,  Ibid.  137,  439- 

441. 
1905    Posternak,  S.:  Sur  la  composition  chimique  et  la  signification  des  grains 

d'aleurone,  Ibid.  140,  322-324. 

1905  Pouchet,  G.,  and  J.  Chevalier:  Action  des  composes  phosphores  sur  la 

circulation,  Bui.  gener.  de  therapeut.     150,  915-921. 

1906  Pouchet,  G.,  and  J.  Chevalier:  Action  du  phosphore  et  des  composes  phos- 

phores organiques  sur  la  circulation,  Ibid.  151,  307-312. 
1905    Power,  Frederick  B.,  and  Frank  Tutin:  The  Relation  between  Natural 
and  Synthetical  Glycerylphosphoric  Acids,  Proc.  of  the  Chem.  Soc.  21, 
72,73;  also  Jour,  of  the  Chem.  Soc.  87  (1905),  249-257. 

1907  Preti,  Luigi:  Ueber  die  spontane  Ausscheidung  einer  Caseinverbindung 

aus  Milch,  Zeit.  physiol.  Chem.  53,  419-426. 


674  OHIO  EXPEKIMENT  STATION:  TECHNICAL  BUL.  5 

1891  Preysz,  Kornel:  (Wie  hat  man  auf  die  Ausscheidung  der  Phosphor- 
saure beziigliche  Versuche  anzustellen  ? ) ,  Magyar  orvosi  archivum  1891, 
50;  through  Jahresb.  u.  d.  Fortschr.  d.  Thierchem.  21  (1891),  252,  353. 

1892-3  Preysz,  Kornel:  Einfluss  der  Muskelarbeit  und  der  geistigen  thatigkeit 
auf  die  Ausscheidung  der  Phosphorsaure  beim  Menschen,  Ungar. 
Arch.  Med.  1,  38-43. 

1871  Pribram,  Richard:  Eine  neue  Methode  zur  Bestimmung  des  Kalkes  und 
der  Phosphorsaure  im  Blutserum,  Ber.  u.  Verhandl.  d.  k.  sachs.  Gesell. 
d.  Wissensch.  23,  279-284.      - 

1898  Prbscher,  Fr.:  Die  Beziehungen  der  Wachsthumsgeschwindigkeit  des 
Sauglings  zur  Zusammensetzung  der  Milch  bei  verschiedenen  Sauge- 
thieren,  Zeit.  physiol.  Chem.  24,  285-302. 

1910  Proskauer,  Felix:  Ueber  den  Erdalkaligehalt    des    Sauglingsblutes    bei 

Ernahrungsstorungen,  Arch.  Kinderheilk.  54,  58-64. 
(1906)  Pruszynski,  J.,  and  J.  Siemienski:  (Untersuchungen  iiber  die  Zusammen- 
setzung der  menschlichen  Galle),  Gazeta  lekarska  26,  225-232;  through 
Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.    36  (1906),  467,  468. 

1911  Pusanow,  Helene:  Bestimmungen  von  Ca,  Mg,  S,  P,  and  CI  im  Blute  von 

Gesunden  und  Kranken,  Diss.,  Zurich.      19  pp.      Through  Zentralbl. 

Biochem.  u.  Biophys.  13  (1912),  101. 
1900    Rachford,  B.  K.:  Pancreatic  Digestion  of  Casein,  Archives  of  Pediat.  17, 

413-426. 
1913    Rakoczy,  A.:  Weiteres  iiber  die  Pepsin-Chymosin-Frage,  Zeit.  physiol. 

Chem.  84,  329-353. 
1887    Ralfe,  Charles  Henry:  Phosphatic  Diabetes,  Lancet  1887, 1,  411-413;  462- 

464. 
1910     Ramacci,  A.:  Sul  quantitative  in  Ca  nel  latte  di  donna,  La  Pediatria  18 

(1910),  665-669:  through  Zentralbl.  Biochem.  u.  Biophys.  11,  882. 
1886    Raske,  K.:  Zur  chemischen  Kenntniss  des  Embryo,  Zeit.  physiol.  Chem. 

10,  336-345. 

1884  Raspopoff,  W.  A.:  Phosphati  v  mochie  bolnich  pri  stradanii  kostei  (Die 

Phosphate  im  Ham  bei  Knochenleiden),  Wratsch  1884,  No.  29,  30  and 
33,  5,  480-511;  through  Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  14 
(1884),  472. 

1885  Raspopoff,  Vladimir:  Ob  usvoenii  i  videlenii  azota  i  fosfornoi  kisloti  pri 

bolezniach  kostei  u  cheloveka  (Application  and  Doses  of  Nitrogen  and 

Phosphoric  Acid  in  Diseases  of  the     Bones),  Diss.,     St.     Petersburg. 

Ill +  68  pp. 
1905-6  Rasquin,  M. :  Les  matieres  phophatees  dans  l'alimentation  des  jeunes 

animaux,  Rapports  II  Congr.  internat.  de     l'aliment.     rationelle     du 

betail,  62-68. 
1912    Rather,  J.  B.:  The  Forms  of  Phosphorus  in  Cottonseed  Meal,  Texas  Agr. 

Exp.  Sta.  Bui.  146,  3-16. 
1913a  Rather,  J.  B.:  The  Phosphorus  Compounds  of  Cotton  Seed  Meal  and 

Wheat  Bran,  Texas  Agr.  Exp.  Sta.  Bui.  No.  156.      18  pp. 
1913b  Rather,  J.  B.:  Phytic  Acid  in  Cottonseed  Meal  and  Wheat  Bran,  Jour. 

Amer.  Chem.  Soc.  35,  890-895. 
1893     Raudnitz,  R.  W. :  Ueber  die  Resorption     alkalischer    Erden    im    Ver- 

dauungstract,  Exp.  Arch.  31,  343. 
1903     Raudnitz,  R.  W.:  Bestandteile,  Eigenschaften  und  Veranderungen  der 

Milch,  Ergebnisse  d.  Physiol.  2,  1.  Abt.s  193-325o      669  refs. 


PHOSPHORUS  METABOLISM  675 

1858  von  Recklinghausen,  F.:  Die  mineralischen  Bestandtheile  junger  Mensch- 
enknochen,  Arch.  path.  Anat.  u.  Physiol.  14,  466-476. 

1891  von  Recklinghausen,  F.:  Die  fibrose  oder  deformirende  Ostitis,  die  Osteo- 
malacic und  die  osteoplastische  Carcinose  in  ihren  gegenseitigen  Bezie- 
hungen,  Festschr.  d.  Assistenten  R.  Virchow's,  1891.      89  pp. 

1879  Regnard,  P. :  Sur  la  composition  chimique  des  os  dans  l'arthropathie  des 
ataxiques,  Compt.  rend.  Acad,  des  sci.  89,  1041,  1042. 

1908  Reh,  Alfred:  Ueber  die  Polypeptidphosphorsaure  (Paranucleinsaure)  des 
Kaseins,  Beitrage  z.  chem.  Physiol,  u.  Path.  11,  1-18. 

(1911)  Reicher,  K.:  Zur  Kenntnis  des  Fett-  und  Lipoidstoffwechsels,  Verhandl. 
d.  Kongr.  f.  innere  Med.  28,  327-330;  through  Jahresb.  u.  d.  Fortschr. 
d.  Thierchem.  41   (1911),  107. 

1904  Reiss,  Emil:  Eine  Beziehung  des  Lecithins  zu  Fermenten,  Berlin,  klin. 

Wochenschr.  41,  1169-1171. 

1894  Rem-Picci,  G.,  and  G.  Bernasconi:  Sull  eliminazione  dei  fosfati  per  le 

urine  nelle  febbri  malariche,  II  Policlinico,  Apr.  1894;  through  Jahresb. 
u.  d.  Fortschr.  d.  Thierchem.  24  (1894),  574-581. 
1913     Renall,  Montague  H. :  Ueber  den  stickstoffhaltigen  Bestandteil  des  Keph- 
alins,  Biochem.  Zeit.  55,  296-300. 

1911  Rengniez,  P.:  Le  phosphore  dans  les  farines,  These  de  pharmacy,  Paris, 

1911;  through  Revue  scient.  49  (1911),  176,  177. 

1905  Renner :  Kunstliche  Hyperleukocytose  als  Mittel  zur  Erhohung  der  Wider- 

standskraft  des  Korpers  gegen  operative  Infektionen.  Wirkung 
subkutaner  Hefenuklei'nsaureinjektionen  auf  den  menschlichen  Organ- 
ismus,  Mittheil.  aus  d.  Grenzgebiet.  d.  Med.  u.  Chir.,  Jena,  15,  89-150. 

1910  Renshaw,  R.  R.,  and  K.  N.  Atkins:  Bactericidal  Properties  of  Lecithins 
and  Choline  Salts,  Jour.  Amer.  Chem.  Soc.  32,  130-132. 

1904  Renvall,  G.:  Zur  Kenntniss  des  Phosphor-,  Calcium-  und  Magnesium- 
umsatzes  beim  erwachsenen  Menschen,  Skand.  Arch.  Physiol.  16,  94- 
138. 

1913     Ribbert,  Hugo:  Beitrag  zur  Rachitis,  Deut.  med.  Wochenschr.  39,  I,  8-10. 

1900  Richet,  Charles:  Du  serum  musculaire,  Compt.  rend.  Acad,  des  sci.  131, 

1314-1316. 

1901  Richmond,  H.  Droop:  The  Composition  of  Milk,  The  Analyst  26,  310-318. 

1912  Riedel,  A.  G.  J.  D.:  Zur  Kenntnis  des  Eigelblecithins,  Riedel's  Ber.,  Mar. 

1912,  24-33;  through  Zentralbl.  Biochem.  u.  Biophys.  13  (1912),  562, 
563. 

1868  Riesell,  Albert:  Ueber  die  Phosphorsaure-Ausscheidung  im  Ham  bei 
Einnahme  von  kohlensaurem  Kalk,  Hoppe-Seyler's  Med.  chem.  Unters. 
319-324. 

1910  Rising,  Adolf:  Bidrag  till  kannedomen  omde  i  vara  narungsmedel  fdre- 
kommande  fosforforeningama  (Phosphorus  Compounds  in  Food  Ma- 
terials), Svensk.  Kemisk  Tidskr.  22,  143-150;  through  Chem.  Abs.  5 
(1911),  2402. 

1895  Robert:  Rachitis  und  Osteomalacia,  Ber.  u.  d.  Veterinarwesen    im    K. 

Sachsen  40,  91. 

1906  Robert,  H.,  and  J.  Parisot:  Les  phosphates  urinairies  dans  la  paralysie 

agitante,  Compt.  rend.  Soc.  de  biol.  60,  1084,  1085. 
1879     Roberts,  William:  Note  on  the  Existence  of  a  Milk-Curdling  Ferment  in 

the  Pancreas,  Proc.  Roy.  Soc,  London,  29,  157. 
1881     Roberts,  William:  On  the  Estimation  of  the  Amylolytic  and  Proteolytic 

Activity  of  Pancreatic  Extracts,  Ibid.  32,  145-161. 


676  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1906-7  Robertson,  T.  Brailsford:  Studies  in  the  Chemistry  of  the  Ion-Proteid 
Compounds.  IV.  On  Some  Chemical  Properties  of  Casein  and  their 
Possible  Relation  to  the  Chemical  Behavior  of  Other  Protein  Bodies, 
with  Especial  Reference  to  Hydrolysis  of  Casein  by  Trypsin,  Jour. 
Biol.  Chem.  2,  317-383. 

1907  Robertson,  T.  Brailsford:  Note  on  the  Synthesis  of  a  Protein  through  the 

Action  of  Pepsin,  Ibid.  3,  95-99. 

1908  Robertson,  T.  Brailsford:  On  the  Influence  of  Temperature  upon  the  Sol- 

ubility of  Casein  in  Alkaline  Solutions,  Ibid.  5,  147-154. 

1909a  Robertson,  T.  Brailsford:  On  the  Synthesis  of  Paranuclein  through  the 
Agency  of  Pepsin  and  the  Chemical  Mechanism  of  the  Hydrolysis  and 
Synthesis  of  Proteins  through  the  Agency  of  Enzymes,  Ibid.  5,  493-523. 

1909b  Robertson,  T.  Brailsford:  On  the  Nature  of  the  Chemical  Mechanism 
which  Maintains  the  Neutrality  of  the  Tissues  and  Tissue-fluids,  Ibid. 
6,  313-320. 

1910a  Robertson,  T.  Brailsford:  Concerning  the  Relative  Magnitude  of  the 
Parts  Played  by  the  Proteins  and  by  the  Bi-carbonates  in  the  Main-? 
tenance  of  the  Neutrality  of  the  Blood.  Ibid.  7,  351-357. 

1910b  Robertson,  T.  Brailsford:  On  the  Refractive  Indices  of  Solutions  of  Cer- 
tain Proteins.      II.  The  Paranucleins,  Ibid.  8,  287-295. 

1911  Robertson,  T.  Brailsford,  and  H.  C.  Biddle:  On  the  Composition  of  Cer- 
tain Substances  Produced  by  the  Action  of  Pepsin  upon  the  Products 
of  Complete  Peptic  Hydrolysis  of  Casein,  Ibid.  9,  295-302. 

1908  Robertson,  T.  Brailsford,  and  C.  L.  A.  Schmidt:  On  the  Part  Played  by 
the  Alkali  in  the  Hydrolysis  of  Proteins  by  Trypsin,  Ibid.  5,  31-48. 

1913  Robertson,  T.  B.,  and  Hardolph  Wasteneys:  On  the  Changes  in  Lecithin 
Content  which  Accompany  the  Development  of  Sea-urchin  Eggs,  Arch. 
Entwickelungsmechanik  d.  Organismen  37,  485-496. 

1895  Robin,  Albert:  Les  glycerophosphates;  Action  sur  la  nutrition,  indica- 
tions therapeutiques,  mode  d'administration,  Bui.  gener.  de  therapeut. 
128,  385-400;  433-444. 

1911  Robin,  Albert:  Beitrag  zur -chemischen  Zusammensetzung  der  carcino- 
matosen  Leber,  Zentralbl.  f.  d.  Physiol,  u.  Path.  d.  Stoffw.,  N.  F.  6, 
577-583;  through  Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  41  (1911), 
344. 

1913a  Robin,  Albert:  Sur  la  mineralisation  comparee  des  regions  cancerisees  du 
foie  et  des  regions  relativement  saines,  Compt.  rend.  Acad,  des  sci. 
156,  334-336. 

1913b  Robin,  Albert :  Recherches  sur  les  variations  de  l'acide  phosphorique  dans 
l'urine  et  le  foie  des  cancereux,  Ibid.  156,  2018-2020. 

1895  Rockwood,  C«  W.:  Ueber  das  Vorkommen  der  Fleischsaure  im  Harne, 
Arch.  Anat.  u.  Physiol.,  physiol.  Abt.,  1-4. 

1898  Rohmann,  F.:  Stoffwechselversuche  mit  phosphorhaltigen  und  phosphor- 
freien  Eiweisskbrpern,  Berlin,  klin.  Wochenschr.  35,  789-791. 

1902  Rohmann,  F.:  Ueber  kiinstliche  Ernahrung.  Vortrag.  Klin.-ther.  Woch- 
enschr. 9,  1306-1311. 

1907  Rohmann,  F.:  Ueber  kiinstliche  Ernahrung,  Deut.  med.  Wochenschr.  33, 

1568. 

1908  Rohmann,  F.:  Ueber  kiinstliche  Ernahrung  von  Mausen,  Allgem.  med. 

Central-Zeitung  77,  129. 


PHOSPHORUS  METABOLISM  677 

I860    Roll,  M.  F.;  Die  Knochenweiche  (Rachitis),  Lehrbuch  der  Path.  u.  Ther. 

d.  Hausthiere,  2d.  edit.,  p.  85. 
1897     Roeske,  Georg:  Ueber  den  Verlauf     der     Phosphorsaure-Ausscheidung 

beim  Menschen,  Inaug.  Diss.,  Greifswald,  44  pp. 

1908  Roger,  H.:  Sur  le  role  des  phosphates  dans  la  saccharification  salivaire, 

Compt.  rend.  Soc.  de  biol.  65,  374,  375. 

1909  Rogers,  L.  A.:  (Gegorene  Milch),  Report    of    the    Bureau    of    Animal 

Industry  1909,  133-161;  through  Jahresb.  ii.  d.  Fortschr.  d.  Thierchem. 
41  (1911),  185. 

1910  Rogozinski,  F.:  Beitrage  zur  Kenntnis  des    Phosphorstoffwechsels    im 

tierischen  Organismus,  Bui.  internat.  de  l'Acad.  des  sci.  de  Cracovie, 

B,  1910,  2,  260-312. 
1909    Rohde,  Alice,  and  Walter  Jones:  The  Purin  Ferments  of  the  Rat,  Jour. 

Biol.  Chem.  7,  237-247. 
1866    Roloff,  F.:  Ueber  Osteomalacia  und  Rachitis,    Arch.    path.    Anat.    u. 

Physiol.  37,  433-503. 
1869    Roloff,  F.:  Ueber  .Osteomalacie,  Ibid.  46,  305-316. 
1875a  Roloff,  F.:  Ueber  die  Ursache  der  Rhachitis,  Zeit.  landwirtsch.  Central- 

Vereins  d.  Prov.  Sachsen  32,  261-263. 
1875b  Roloff,  F.:  Ueber  Osteomalacie  und  Rachitis.      I.  Arch,  prakt.  Tierheilk. 

1,  189-220. 
1879    Roloff,  F.:  Ueber  Osteomalacie  und  Rachitis.      II.  Ibid.  5,  152-163. 
1887     Romeyn,  D. :  Ondersoekingen  over  den  invloed  van  alcohol  op  den  Mensch, 

Akad.  Prolfschrift,  Amsterdam,  1887;  Munch,  med.  Wochenschr.  1887, 

No.  33;  through  Jahresb.  ii.  d.  Fortschr.  d.    Thierchem.    17     (1887), 

400-403. 
1846    Ronalds,  Edmund:  Remarks  on  the  Extractive  Material  of  Urine,  and  on 

the  Excretion  of  Sulphur  and  Phosphorus  by  the  Kidneys  in  an  Unox- 

idized  State,  Phil.  Trans,  of  the  Roy.  Soc.  of  London  136  (1846),  461- 

464;  also  Phil.  Mag.  [3]  30  (1847),  253;  also  Jour.  f.  prakt.  Chem.  41 

(1847),  185-188. 
1895     Roos,  E.:  Ueber  die  Einwirkung  der  Schilddruse  auf  den  Stoffwechsel 

nebst  Vorversuchen  iiber  die  Art  der  Wirksamen  Substanz  in  derselben, 

Zeit.  physiol.  Chem.  21,  19-41. 
1911     Rose,  Anton  Richard:  The  Toxicity  of  Phytin,  Rept.  Amer.  Chem.  Soc. 

for  1911,  Science  35  (1912),  393. 
1912a  Rose,  Anton  Richard:  A  Study  of  the  Metabolism  and  Physiological  Ef- 
fects of  Certain  Phosphorus  Compounds  with  Milch  Cows.      II.  N.  Y. 

Agr.  Exp.  Sta.,  Tech.  Bui.  No.  20. 
1912b  Rose,  Anton  Richard:  A  Resume  of  the  Literature  on  Inosite-Phosphoric 
Acid,  with  Special  Reference  to  the  Relation  of  that  Substance  to 
Plants,  Biochemical  Bui.  2,  21-49. 

1911  Rose,  A.  R.,  and  J.  T.  Cusick:  The  Influence  of  Phosphorus  Compounds 

on  the  Yield  .and  Composition  of  Goat's  Milk,  Rept.  Amer.  Chem.  Soc. 
for  1911,  Science  35  (1912),  393. 
1849     Rose,  Heinrich :  Ueber  die  unorganischen  Bestandtheile  in  den  organisch- 
en  Korpern,  Poggendorff's  Annalen  d.  Phys.  u.  Chem.  76  (16  of  ser.  3), 
305-404. 

1912  Rosenbloom,  Jacob:  A  Quantitative  Study  of  the  Lipins  of  Bile  Obtained 

from  a  Patient  with  a  Biliary  Fistula,  Proc.   Columbia  Biochem.  Assoc. 
Biochemical  Bui.  2,  182. 


678  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1913a  Rosenbloom,  Jacob:  The  Biochemistry  of  the  Female  Genitalia.  II.  The 
Lipins  of  the  Ovary  and  Corpus  Luteum  of  the  Pregnant  and  Non- 
pregnant Cow,  Jour.  Biol.  Chem.  13,  511,  512. 

1913b  Rosenbloom,  Jacob:  A  Quantitative  Chemical  Analysis  of  Human  Bile, 
Ibid.  14,  241-243. 

1909  Rosenheim,  Otto:  Proposals  for  the  Nomenclature  of  the  Lipoids,  Bio- 

chemical Jour.  4,  331-336. 

1907  Rosenheim,  Otto,  and  M.  Christine  Tebb:  The  Non-existence  of  "Pro- 
tagon"  as  a  Definite  Chemical  Compound,  Jour,  of  Physiol.  36,  1-16. 

1908a  Rosenheim,  Otto,  and  M.  Christine  Tebb:  The  Optical  Activity  of  So- 
called  "Protagon,"  Ibid.  37,  341-347 

1908b  Rosenheim,  Otto,  and  M.  Christine  Tebb:  On  a  New  Physical  Phenom- 
enon Observed  in  Connection  with  the  Optical  Activity  of  So-called 
"Protagon,"  Ibid.  37,  348-354. 

1908c  Rosenheim,  Otto,  and  M.  Christine  Tebb :  On  So-called  "Protagon,"  Quart. 
Jour,  of  Exp.  Physiol.  1,  297-304. 

1909a  Rosenheim,  Otto,  and  M.  Christine  Tebb:  The  Non-Existence  of  "Pro- 
tagon" as  a  Definite  Chemical  Compound,  Ibid.  2,  317-333. 

1909b  Rosenheim,  Otto,  and  M.  Christine  Tebb:  On  the  Lipoids  of  the  Adrenals. 
(Preliminary  communication.)  Proc.  Physiolog.  Soc,  Jour,  of 
Physiol.  38,  LIV-LVI. 

1909c  Rosenheim,  Otto,  and  M.  Christine  Tebb:  The  Lipoids  of  the  Brain.  Part 
I.  Sphingomyelin,  Ibid  38,  LI-LII. 

1910  Rosenheim,  Otto,  and  M.  Christine  Tebb:  Die  Nicht-Existenz  des  sogen- 

annten  "Protagons"  im  Gehirn,  Biochem.  Zeit.  25,  151-160. 

1901  de  Rothschild,  H.,  and  L.  Netter:    Apropos  des  quantites  de  lait  qu'il 

convient  de  donner  dans  l'allaitement  artificiel  et  de  leurs  rapports 
avec  les  echanges  nutritifs  chez  le  nourrisson,  Compt.  rend.  Soc.  de 
Biol.  53,  658-661. 
1910    Rotky,  Hans:  Beitrage  zur  Pathologie  des  Nucleinstoffwechsels,  Deut. 
Arch.  klin.  Med.  98,  540-582. 

1902  Rotondi,  Giorgio:  Sulla  digestione  del  caseinogeno  di  donna  e  di  mucca, 

Lo  Sperimentale  56,  68-83;  through  Jahresb.  ii.  d.  Fortschr.  d.  Thier- 
chem.  32  (1902),  999-1000. 
1912     Rubinraut,  A.:     Stoffwechselversuche    bei    Akromegalie,    Inaug.     Diss. 
Zurich.  19  pp. 

1903  Rubow,  V.:  Unders0gelser  over  normale  og  fedtdegenererede  Hjaerter 

som  Bidrag  til  Hjaertemusklens  physiologiske  Kemi  og  til  Fedtdegen- 
erationens  (Researches  on  normal  and  fatty  degenerated  hearts  as  a 
contribution  to  the  physiological  chemistry  of  the  heart  muscles  and 
to  the  pathogenesis  of  fatty  degeneration),  K0benh.  128  pp. 

1905  Rubow,  V.:  Ueber  den  Lecithingehalt  des  Herzens  und  der  Nieren  unter 
normalen  Verhaltnissen,  im  Hungerzustande  und  bei  der  fettigen  De- 
generation, Arch.  exp.  Path.  u.  Pharm.  52,  173-204. 

1893a  Riidel,  G.:  Ueber  die  Resorption  und  Ausscheidung  des  Kalkes,  Ibid.  33, 
79-89. 

1893b  Riidel,  G.:  Ueber  die  Resorption  und  Ausscheidung  der  Kalksalzen  bei 
rachitischen  Kindern,  Ibid.  33,  90-100. 

1900  Rumpf,  Th.,  and  O.  Schumm:  Ueber  den  Stoffwechsel  eines  Vegetariers, 
Zeit.  Biol.  39  (N.  F.  21),  153-158. 


PHOSPHORUS  METABOLISM  679 

1895    Ruppel,  W.  G. :  Zur  Kenntniss  des  Protagons,  Ibid.  31,  86-100. 

1912    Ryan,  L.  A.,  and  Edward  B.  Meigs:  The  Chemical  Constituents  of  the 

Ash  of  Smooth  Muscle,  Proc.  Amer.  Soc.  Biol.  Chem.,  Jour.  Biol.  Chem. 

11  (1912),  XXV;  also  Proc.  Amer.  Physiol.  Soc,  Amer.  Jour.  Physiol. 

29  (1912),  XV,  XVI. 

1907  Saccone,  Giovanni:  Sul  ricambio  del  fosforo  dopo  lesione  od  asportazione 

di  organi  importanti,  Annali  di  med.  navale  13,  I,  573-598. 
1905     Sachs,  Fritz:  Ueber  die  Nuklease,  Zeit.  physiol.  Chem.  46,  337-353. 
1891     Sagnier,  Henri:  L'industrie  laitiere  a  Vichy,  Jour,  de  l'agriculture,  26, 

2,  470-473;  through  Milch-Zeitung  20  (1891),  890. 

1908  Saikai,  Tadasu:  A  Chemical  Study  of  Nonstriated  Mammalian  Muscle, 

Jour.  Biol.  Chem.  4,  483-495. 
1900     Saint- Aubin,  Henri    S.:  Injections  hypodermiques  d'huile  de  vaseline  et 

de  jaune  d'oeuf  (Medication  stimulante),  These  de  Paris.       41  pp. 
(1907)  St.  Bondzynski  and  A.  Gonka:  (Ein  Beitrag  zur  Kenntnis  der  organisch- 

en  Grundsubstanz  der  Zahne),     Sprawozdanie    X    zjardu    lekarzy    I 

przyrodnikow  polskich  S.  105,  vorl.  Mitteil.;  through  Jahresb.  ii.  d. 

Fortschr.  d.  Thierchem.  37  (1907),  478. 
1912     Sakaki,  C:  Ueber  einige  Phosphatide  in  der  Placenta,  Mitt.  Med.  Ges. 

Tokio  26,  No.  22, 1. 
1913a  Sakaki,  C:  Ueber  einige  Phosphatide  aus  der  menschlichen  Placenta.  I. 

Biochem.  Zeit.  49,  317-325. 
1913b  Sakaki,  C:  Ueber  einige  Phosphatide  aus  der  menschlichen  Placenta.  II. 

Ibid.  49,  326-332. 
1913c  Sakaki,  C:  Ueber  einige  Phosphatide  aus  der  menschlichen  Placenta.  III. 

Ibid.  54, 1-4. 
1913d  Sakaki,  C:  Ueber  die  Phosphorverteilung  in  der  Placenta,  Ibid.  54,  5-10. 
1910     Salant,  William,  and  F.  C.  Hinkel:  The  Influence  of  Alcohol  on  the  Com- 
position of  Urine.       I.  Jour,  of  Pharmacol,  and  Exper.  Therapeut.  1, 

493-517. 
1885     Salkowski,  E.:  Zur  Kenntnis  des  Pferdeharns,  Zeit.  physiol.  Chem.  9,  241- 

245. 
1893a  Salkowski,  E.:  Ueber  den  Verbleib  des  Phosphors  bei  der  Verdauung  des 

Caseins,  Centralbl.  med.  Wissensch.  31,  385,  386. 
1893b  Salkowski,  E.:  Ueber  das  Verhalten  des  Caseins  bei  der  Magenverdau- 

ung  und  die  Verseifung  der  Fette,  Ibid.  31,  467-469. 
1896a  Salkowski,  E. :  Ueber  das  Verhalten  des  Caseins  zu  Pepsinsalzsaure,  Arch. 

ges.  Physiol.  63,  401-422. 
1896b  Salkowski,  E.:  Ueber  die    Anwendung    eines    neuen    Caseinpraparates 

"Eucasein"  zu  Ernahrungszwecken,  Deut.  med.  Wochenschr.,  225-229. 
1899     Salkowski,  E.:  Kleinere  Mittheilungen.      1.  Ueber  das  erste  Produkt  der 

Verdauung  des  Caseins  durch  Pepsinsalzsaure,  Zeit.  physiol.  Chem. 

27,  297-302. 
1901     Salkowski,  E. :  Ueber  die  Paranucleinsaure  aus  Casein.      I.  Ibid.  32,  245- 

267.     • 
1904     Salkowski,  E.:  Zur  Kenntnis  des  Harns  und  des  Stoffwechsels  der  Her- 

bivoren,  Ibid.  42,  212-250. 
1909a  Salkowski,  E.:  Ueber  das  Ferratin  Schmiedebergs,  Ibid.  58,  282-289. 
1909b  Salkowski,  E.:  Ueber  die  Bindung  des  Eisens    im    Nucleoproteid    der 

Leber,  Ibid.  59,  19-21. 


680  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1913a  Salkowski,  E.:  Ueber  das  Verhalten  des  jodparanucleinsauren  Eisens  im 

Organismus,  Biochem.  Zeit.  49,  152-164. 
1913b  Salkowski,  E.:  1st  es  moglich,  den  Gehalt  des  Gehirns  an  Phosphatidyl 

zu  steigern?      Ibid.  51,  407-422. 
1894-5  Salkowski,  E.,  and  Martin  Hahn:  Ueber  das  Verhalten  des  Phosphors  im 

Casein  bei  der  Pepsinverdauung,  Arch.  ges.  Physiol.  59,  225-250. 
1878     Salomon,  Georg:  Ueber  die  Verbreitung  und  Entstehung  von  Hypoxan- 

thin  und  Milchsaure  im  thierischen  Organismus,  Zeit.  physiol.  Chem. 

2,  65-95. 
1907a  Sammis,  J.  L.:  The  Chemistry  of  Milk  Curdling,  Twenty-fourth     Ann. 

Kept.  Wis.  Agr.  Exp.  Sta.,  171-189. 
1907b  Sammis,  J.  L.:  The  Coagulation  of  Fresh  and  Alkaline  Milks,  Ibid.  190- 

206. 

1909  Sammis,  J.  L.,  and  E.  B.  Hart:  The  Relation  of  Different  Acids  to  the 

Precipitation  of  Casein  and  to  the  Solubility  of  Cheese  Curds  in  Salt 
Solution,  Jour.  Biol.  Chem.  6,  181-187. 

1895  Sandmeyer,  W.:  Ueber  die  Ausnutzung  des  Paranucleins  im  tierischen 

Organismus,  Zeit.  physiol.  Chem.  21,  87-89. 
1912     Sani,  G.:  Azione  del  fosfato  monocalcico  nella  conservazione  dei  foraggi 

verdi,  Atti.  Accad.  Lincei  21,  II,  108-112. 
1894     Sanson,  Andre:  Sur  l'enrichissenient  du  lait  en  phosphates,  Compt.  rend. 

Soc.  de  biol.  46,  154,  155. 

1896  Sanson,  Andre:  Sur  l'assimilabilite  des  glycerophosphates,  Ibid.  48,  685- 

687. 

1912  Santonocetto,  Vittorugo:  La  fitina  nel  ricambio  organico  azotato  fisio- 

logico,  Bullett.  delle  sci.  med.  di  Bologna,  223-232;  through  Zentralbl. 
Biochem.  u.  Biophys.  14  (1912-13),  476. 
(1911)  Sarvonat,  F.,  and  Gentz:  (Ueber  den  Verlauf  der  Phosphorsaureausschei- 
dung  im  Harn),  Compt.  rend.  Soc.  de  biol.  70,  629;  through  Jahresb.  ii. 
d.  Fortschr.  d.  Thierchem.  41  (1911),  468. 

1910  Sarvonat,  F.,  and  Rebattu,  J.:  Action  de  la  tuberculose  sur  la  mineral- 

isation du  cobaye,  Jour,  de  physiol.  et  de  pathol.  gener.  12,  934-941; 
also  Compt.  rend.  Soc.  de  biol.  69  (1910),  127-129. 

1909  Sato,  T.:  Beitrag  zur  Kenntniss  des  Nucleoproteids  der  Milz,  Biochem. 

Zeit.  22,  489-494. 

1907  Satta,  G.:  Sulla  distribuzione  del  fosforo  negli  organi  e  sulla  decompos- 

izione  dei  composti  fosforati  organici.       Parte  prima.       Archivio  sci. 
med.  Torino  31,  407-412. 

1908  Satta,  G.:  Sur  la  distribution  du  phosphore  dans  les  organes  et  sur  la 

decomposition  des  composes  phosphores  organiques,  Archives  ital.  d. 
biol.  49,  375-380. 

1910  Satta,  G.,  and  G.  M.  Fasiani:  Influenza  dei  lipoidi  sull'  autolisi  del  fegato, 

Giornale  d.  reale  Accad.  di  med.  di  Torino  73  (16  of  ser.  4),  285-288. 

1913  Satta,  G.,  and  G.  Gastaldi:  Sul  ricambio  della  lecitina  introdotta  per 

bocca,  Archivio  di  farmacol.  speriment.  15,  402-406. 
1902     Sauerbruch:  Ein  Beitrag  zum  Stoffwechsel  des  Kalkes  und  der  Phos- 

phorsaure  bei  infantiler  Osteomalacic,  Inaug.  Diss.,  Leipzig.      51  pp. 
1910     Sauerland,  F.:  Ueber  den  Eisengehalt  der  echten  Nucleinsaure,  Zeit. 

physiol.  Chem.  64,  16-20. 


PHOSPHORUS  METABOLISM  681 

1911     Sawadski,  J.  W.:  (Klinische  Beobachtung  iiber  den  Phosphor  des  Bultes), 

Russki  Wratsch,  Nr.  54;  through  Jahresb.  ii.  d.  Fortschr.  d.  Thierchem. 

41  (1911),  136. 
1908    Scaffidi,  Vittorio:  Ueber  die  Verteilung  des  Eisens  in  der  Leber,  Zeit. 

physiol.  Chem.  54,  448-460. 
1909a  Scaffidi,  Vittorio:  Ueber  das  Nucleoproteid  der  Schweinsleber,  Ibid.  58, 

272-281. 
1909b  Scaffidi,  Vittorio:  (Purine  Excretion  in  Animals  Treated  with  Nucleic 

Acid),  Lo  Sperimentale  63,  243-253;  through  Chem.  Abs.  3   (1909), 

2990. 
1910-11  Scaffidi,  Vittorio:  Sulle  modificazioni  del  ricambio  purinico  nella  fatica 

in  alta  montagna,  Internat.  Beitrage  z.  Path.  u.  Ther.  der  Ernahrungs- 

storungen  2,  145-183. 

1907  Schabad,  J.  A.:  (Der  Einfluss  des  Phosphors  auf  den  Kalkstoffwechsel 

bei  rachitischen  und  gesunden  Kindern),  Aerzte  Zeitung  1907,  No.  13 
(Russian);  through  Jahresb.  ii.  d.  Fortsch.  d.  Thierchem.  37  (1907), 
661. 

1908  Schabad,  J.  A.:  Der  Phosphorlebertran  in  der  Therapie  der  Rhachitis. 

Sein  Einfluss  auf  den  Kalkstoffwechsel,  Therapie  der  Gegenwart  49, 
260,  261. 

1909a  Schabad,  J.  A.:  Der, Phosphor  in  der  Therapie  der  Rachitis.  Der  Ein- 
fluss des  Phosphors  auf  den  Kalkstoffwechsel  bei  rachitischen  und  ge- 
sunden Kindern,  Zeit.  klin.  Med.  67,  454-494. 

1909b  Schabad,  J.  A.:  Die  Behandlung  der  Rachitis  mit  Lebertran,  Phosphor 
und  Kalk.  Ihr  Einfluss  auf  den  Kalk-  und  Phosphorstoffwechsel  bei 
Rachitis,  Ibid.  68,  94-110. 

1909c  Schabad,  J.  A.:  Zur  Bedeutung  des  Kalkes  in  der  Pathologie  der  Rach- 
itis. Abhandlung  I.  Der  Mineralgehalt  gesunder  und  rachitischer 
Knochen,  Arch.  Kinderheilk.  52,  47-67. 

1909d  Schabad,  J.  A. :  Zur  Bedeutung  des  Kalkes  in  der  Pathologie  der  Rachitis. 
Abhandlung  II.  Der  physiologische  Kalkbedarf  und  Rachitis  infolge 
von  unbefriedigtem  Kalkbedarf,  Ibid.  52,  68-106. 

1909e  Schabad,  J.  A.:  Der  Kalk  in  der  Pathologie  der  Rachitis,  Berlin,  klin. 
Wochenschr.  46,  823-826;  923-925. 

1910a  Schabad,  J.  A.:  Die  gleichzeitige  Verabreichung  von  Phosphorlebertran 
mit  einem  Kalksalze  bei  Rachitis,  Jahrb.  Kinderheilk.     72,  1-17. 

1910b  Schabad,  J.  A.:  Phosphor,  Lebertran  und  Sesamol  in  der  Therapie  der 
Rachitis,  Zeit.  klin.  Med.  69,  435-474. 

1910c  Schabad,  J.  A.:  Zur  Bedeutung  des  Kalkes  in  der  Pathologie  der  Rachitis. 
Abhandlung  III.  Der  Kalkstoffwechsel  bei  Rachitis,  Arch.  Kinderheilk. 
53,  380-414. 

1910d  Schabad,  J.  A. :  Zur  Bedeutung  des  Kalkes  in  der  Pathologie  der  Rachitis. 
Abhandlung  IV.  Der  Phosphorstoffwechsel  bei  Rachitis,  Ibid.  54,  83- 
110. 

1910-11  Schabad,  J.  A.:  Der  Kalkstoffwechsel  bei  Tetanie,  Monatsschr.  Kinder- 
heilk. 9  (Originalien  1),  25-35. 

1911a  Schabad,  J.  A.:  Der  Kalkgehalt  der  Frauenmilch.  Zur  Frage  der  un- 
genugenden  Kalkzufuhr  als  Ursache  der  Rachitis,  Jahrb.  Kinderheilk. 
74,  511-535. 


682  OHIO  EXPERIMENT  STATION:  TECHNICAL  *BUL.  5 

(1911b)  Schabad,  J.  A.:  Zwei  Falle  von  sogenannter  "Spatrachitis."  Der  Min- 
eralgehalt  der  Knochen  und  der  Mineralstoffwechsel  im  Vergleich  zu 
der  kindlichen  Rachitis,  Mitt.  a.  d.  Grenzgeb.  d.  Med.  u.  Chirurg.  23, 
82-99;  through  Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  41  (1911),  360. 

1911c  Schabad,  J.  A.:  (Der  Aschegehalt  der  Knochen  und  der  Mineralstoffwech- 
sel im  Vergleich  mit  dem  bei  Rachitis),  Wratschebnaja  Gazeta  1911, 
Nr.  10  u.  11;  through  Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  41  (1911), 
360. 

1911a  Schabad,  J.  A.,  and  R.  F.  Sorochowitsch:  Lipanin  als  Ersatzmittel  des 
Lebertrans  bei  Rhachitis.  Sein  Einfiusz  auf  den  Stoffwechsel,  Monat- 
schr.  Kinderheilk.      9  (Originalien  1),  659-671. 

1911b  Schabad,  J.  A.,  and  R.  F.  Sorochowitsch:  Die  Behandlung  des  Rachitis 
mit  Lebertranemulsionen  (Emulsion  Scott  and  Parke  Davis  mit  Nat- 
rium- und  Calciumhypophosphit  und  Emulsion  mit  essigsaurem  Kalk) 
und  ihre  Einwirkung  auf  den  Stoffwechsel,  Ibid.  10  (Originalien  2),  12- 
34. 

1912  Schabad,  J.  A.,  and  R.  F.  Sorochowitsch :  Zur  Frage  von  Wesen  der  giinst- 
igen  Wirkung  des  Lebertrans  bei  Rhachitis.  1st  das  wirksame 
Agens  des  Lebertrans  ein  Ferment?    Ibid.  11  (Originalien  3),  4-20. 

1889  Schabanowa,  Anna:  Ueber  Phosphorbehandlung  der  Rachitis,  Jahrb. 
Kinderheilk.  29,  392-410. 

1912  Schafer,  E.  A.:  Effects  upon  Growth  and  Metabolism  of  the  Addition  of 
Small  Amounts  of  Ovarian  Tissue,  Pituitary,  and  Thyroid  to  the  Nor- 
mal Dietary  of  White  Rats,  Quart.  Jour,  of  Exp.  Physiol.  5,  203-228. 

1887  Schaffer,  F.:  Ueber  das  Casein  und  die  Wirkung  des  Labfermentes  in  der 
Kuhmilch,  Landwirtsch.  Jahrb.  d.  Schweiz.  1,  43-58. 

1908  Schaumann,  H.:  Beriberi  and  Nucleinphosphorsaure    in    der    Nahrung, 

Arch.  Schiffs-  und  Tropenhygiene  12,  Beiheft  5,  37-57. 

1909  Schaumann,  H.:  Weitere  Beitrage  zur  Aetiologie  der  Beriberi,  Ibid.  13, 

Beiheft  6,  82-90. 

1910  Schaumann,  H.:  Die  Aetiologie  der  Beriberi  unter  Beriicksichtigung  des 

gesamten  Phosphorstoffwechsels,  Ibid.  14,  Beiheft,  8,  385  pp.  294  refs. 
1911a  Schaumann,  H.:  Further  Contributions  to    the    Etiology    of    Beri-beri, 

Trans,  of  the  Soc.  of  Tropical  Med.  5,  59-91. 
1911b  Schaumann,  H.:  Erwiderung  auf  C.  Eijkman,  Polyneuritis  gallinarum 

und  Beriberi,  Arch.  Schiffs-  u.  Tropenhygiene  15,  728-737. 
1912a  Schaumann,  H.:  Ueber  die  Darstellung  und  Wirkungsweise  einer  der  in 

der  Reiskleie  enthaltenen,  gegen  experimentelle     Polyneuritis     wirk- 

samen  Substanzen,  Ibid.  16,  349-361. 
1912b  Schaumann,  H.:  Zu  dem  Problem  der  Beriberiatiologie,  Ibid  16,  825- 

838. 
1912c  Schaumann,  H.:  Weitere  Beitrage  zur  Aetiologie  der  Beriberi,  Ibid.  16, 

Beihefte,  137-170. 
1905  Schenck,  Martin:  Die  bei  der  Selbstverdauung  des  Pankreas  auftretenden 

Nucleinbasen,  Zeit.  physiol.  Chem.  43,  406-409. 
1903     Schenke,  V.:  Untersuchungen  iiber  die  Futtermittel  des  Handels,  veran- 

lasst  1890  auf  Grund  Beschlusse  in  Bernburg  und  Bremen  durch.  den 

Verband     landwirtschaftl.     Versuchs-Stationen  im  Deutschen  Reiche. 

XXXII.  Phosphorsaurer  Kalk  als  Futterbeigabe,  Landwirtsch.  Versuch. 

Stat.  58,  291-312. 


PHOSPHORUS  METABOLISM  683 

1880  Schetelig:  Ueber  die  Herstammung  und  Ausscheidung  des  Kalkes  im  ge- 
sunden  und  kranken  Organismus,  Arch.  path.  Anat.  u.  Physiol.  82, 
437-455. 

1911  Scheunert,  A.,  A.  Schattke  and  E.  Loetsch:  CaO-,  MgO-,  P205-  Gehalt 
von  Heu  und  Hafer,  nach  deren  Verfutterung  Pferde  an  Osteomalacie 
erkrankten,  Biochem.  Zeit.  36,  240-244. 

1897a  Schiff,  Arthur:  Ueber  die  Beeinflussung  des  Stoffwechsels  durch  Hypo- 
physis und  Thyreoideapraparate,  Zeit.  klin.  Med.  32,  Suppl.,  284-290. 

1897b  Schiff,  Arthur:  Hypophysis  und  Thyroidea  in  ihrer  Einwirkung  auf  den 
menschlichen  Stoffwechsel,  Wien.  klin.  Wochenschr.  10,  277-285. 

1901a  Schilling,  Fr.:  Die  Tripelphosphate  im  Stuhle  Ikterischer,  Centralbl. 
Stoffwechsel-  u.  Verdauungskrankh.  2,  391,  392. 

1903  Schittenhelm,  Alfred:  Die  Nucleinbasen  der  Faeces  unter  dem  Einfluss 

anhaltender  Faulnis,  Zeit.  physiol.  Chem.  39,  199-202. 
1904a  Schittenhelm,  Alfred:  Ueber  die  Harnsaurebildung  in  Gewebsausziigen, 

Ibid.  42,  251-258. 
1904b  Schittenhelm,  Alfred :  Ueber  die  Fermente  des  Nucleinstoffwechsels,  Ibid. 

43,  228-239. 
1905a  Schittenhelm,  Alfred:  Ueber  die  Harnsaurebildung  und  die  Harnsaure- 

zersetzung  in  den  Auszugen  der  Rinderorgane.  Ein  weiterer  Beitrag 

zur  Kenntnis  der  Fermente  des  Nuclinstoffwechsels,  Ibid.  45,  121-151. 
1905b  Schittenhelm,  Alfred:  Der  Nucleinstoffwechsel  und  seine  Fermente  bei 

Mensch  und  Tier,  Ibid.  46,  354-370. 

1907  Schittenhelm,  Alfred:  Natur  und  Wesen  der  Gicht,  Med.  Klinik,  Berlin, 

1907,  Beiheft  4,  89-110. 

1908  Schittenhelm,  Alfred:  Ueber  die  Fermente  des  Nucleinstoffwechsels,  Zeit. 

physiol.  Chem.  57,  21-27. 
1909a  Schittenhelm,  Alfred:  Ueber  die  Umsetzung  verfiitterter  Nucleinsaure 

beim  Hunde  unter  normalen  und  pathologischen  Bedingungen,  Ibid. 

62,  80-99. 
1909b  Schittenhelm,  Alfred:  Ueber  die     Fermente     des     Nucleinstoffwechsels 

menschlicher  Organe,  Ibid.  63,  248-268. 
1910     Schittenhelm,  Alfred:  Ueber  den  Nucleinstoffwechsel     des     Schweines, 

Ibid.  66,  53-69. 

1904  Schittenhelm,  A.,   and  Ernst   Bendix:   Ueber  das   Schicksal  der  in  die 

Blutbahn   eingebrachten   Nucleinsaure,    Deut.    med.    Wochenschr.    30, 
1164,  1165. 

1905  Schittenhelm,  A.,  and  Ernst  Bendix:  Vergleichende  Untersuchungen  liber 

die  Wirkung  verschiedener  Nukleinsaure  auf  den  thierischen  Organis- 
mus, Zeit.  exp.  Path.  u.  Ther.  2,  166-178. 

1906  Schittenhelm,  Alfred,  and  Julius  Schmidt:  Ueber    die     Fermente     des 

Nucleinstoffwechsels,  Zeit.  physiol.  Chem.  50,  30-35. 

1907a  Schittenhelm,  Alfred,  and  Julius  Schmidt:  Ablauf  des  Nucleinstoffwech- 
sels in  menschlichen  Organen,  Zeit.  exp.  Path.  u.  Ther.  4,  424-431. 

1907b  Schittenhelm,  Alfred,  and  Julius  Schmidt:  Ablauf  des  Nucleinstoffwech- 
sels in  der  Schweineleber,  Ibid.  4,  432-437. 

1903a  Schittenhelm,  A.,  and  F.  Schroter:  Ueber  die  Spaltung  der  Hefenuclein- 
saure  durch  Bakterien.  I.  Mittheilung,  Zeit.  physiol.  Chem.  39,  203- 
207. 


684  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1903b  Schittenhelm,  A.,  and  F.  Schroter:  Ueber  die  Spaltung  der  Hefenuclein- 
saure  durch  Bakterien.  II.  Mittheilung  and  III.  Mittheilung,  Ibid.  40, 
62-69;  70-80. 

1904  Schittenhelm,  A.,  and  F.  Schroter:  Ueber  die  Spaltung  der  Hefenuclein- 
saure  durch  Bakterien.      IV.  Mittheilung,  Ibid.  41,  284-292. 

(1911)  Schittenhelm,  Alfred,  and  Philip  Seisser:  Ueber  die  Beeinflussung  des 
Stoffwechsels  von  Kaninchen  und  Hund  durch  Zufuhr  von  Nuklein- 
saure,  Harnsaure  und  Allantoin  (gleichzeitig  ein  Beitrag  zur  Frage 
der  Nuklei'nharnsaureverbindungen),  Zeit.  exp.  Path.  u.  Ther.  7,  116- 
133;  through  Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  41  (1911),  477,  478. 

1909  Schittenhelm,  Alfred,  and  Karl  Wiener:  Ueber  die  Vorkommen  und  die 

Bedeutung  von  Allantoin  im  menschlichen  Urin,  Zeit.  physiol.  Chem. 
63,  283-288. 
1912    Schittenhelm,  Alfred,  and  Karl  Wiener:  Ueber  den  Abbau  der  Nuclein- 
saure  durch  Organfermente,  Ibid.  77,  77-85. 

1910  Schkarin,  A.  N.:  Ueber  den  Einfluss  der  Nahrungsart  der  Mutter  auf 

Wachstum  und  Entwicklung  des  Sauglings,  Monatsschr.  Kinderheilk. 
9  (Originalien  1),  65-69. 

1902  Schlagdenhauffen  and  Reeb:  Sur  la  presence  de  la  lecithine  dans  les 
vegetaux,  Compt.  rend.  Acad,  des  sci.  135,  205-208. 

(1911)  Schloss,  Ernst:  Die  chemische  Zusammensetzung  der  Frauenmilch  auf 
Grund  neuer  Analysen,  Monatsschr.  Kinderheilk.  9,  636-640;  Jahresb. 
ii.  d.  Fortschr.  d.  Thierchem.  41  (1911),  206. 

1913a  Schloss,  Ernst:  Zur  Behandlung  der  Rachitis  mit  Lebertran,  Kalk,  und 
Phosphor  auf  Grund  von  Stoffwechselversuchen,  Deut.  med.  Wochen- 
schr.  39,  II,  1505-1508. 

1913b  Schloss,  Ernst:  Zur  Therapie  der  Rachitis.  1.  Mitteilung.  Die  Wirkung 
von  Phosphorlebertran  und  Calcium  aceticum  auf  den  Stoffwechsel 
des  naturlich  ernahrten  rachitischen  Kindes,  Jahrb.  Kinderheilk.  78 
[3rd  ser.  28],  694-722. 

1914a  Schloss,  Ernst:  Zur  Therapie  der  Rachitis.  2.  Mitteilung.  Die  Wirk- 
ung von  Phosphorlebertran,  Calcium  aceticum  und  organischen  Kalk- 
phosphaten  auf  den  Stoffwechsel  des  naturlich  ernahrten  rachitischen 
Kindes,  Ibid.  79  [3rd  ser.  29],  40-65. 

1914b  Schloss,  Ernst:  Zur  Therapie  der  Rachitis.  3.  Mitteilung.  Die  Wirk- 
ung von  organischen  und  anorganischen  Kalkphosphorpraparaten  auf 
den  Stoffwechsel  des  naturlich  ernahrten  rachitischen  Kindes,  Ibid.  79 
[3rd  ser.  29],  194-214. 

1911  Schloss,  Oscar  M.,  and  James  M.  Crawford:  The  Metabolism  of  Nitro- 
gen, Phosphorus,  and  the  Purin  Substances  in  the  New-Born;  with 
special  Reference  to  the  Causation  of  the  Uric  Acid  Infarcts  of  the 
Kidney,  Amer.  Jour.  Diseases  of  Children  1,  203-229. 

1896  Schlossmann,  Arthur :  Ueber  die  Eiweissstoff e  der  Milch  und  die  Method- 

en  ihrer  Trennung,  Zeit.  physiol.  Chem.  22,  197-226. 

1897  Schlossmann,  Arthur:  Ueber  Eselsmilch,  Ibid.  23,  258-264. 

1905a  Schlossmann,  Arthur:  Ueber  die  Bedeutung  des  Phosphors  in  der  Milch 
fur  den  Sauglingsorganismus,  Med.  Klinik  1,  249-251. 

1905b  Schlossmann,  Arthur:  Ueber  Menge,  Art  und  Bedeutung  des  Phosphors 
in  der  Milch  und  iiber  einige  Schicksale  desselben  im  Sauglingsorganis- 
mus, Arch.  Kinderheilk.  40,  1-39. 


PHOSPHORUS  METABOLISM  685 

1904  Schlossmann,  Arthur,  and  Ernst  Moro:  Die  Ernahrung  des  Erwachsenen 
mit  Kuh-  und  mit  Frauenmilch,  Zeit.  Biol.  45,  261-291. 

1847  Schmidt,  Carl:  Knochenerweichung  durch  Milchsaurebildung,  Annalen 
der  Chem.  u.  Pharm.  61,  329-335. 

1882  Schmidt:  (Materialien  zur  Erklarung  der  Eigenschaften  von  Frauen  und 

Kuhmilch),  Russian  Diss.;  through  Centralbl.  Gynakol.  6,  776. 

1883  Schmidt-Miilheim :  Beitrage  zur  Kenntniss  der  Milchsecretion,  Arch.  ges. 

Physiol.  30,  602-620. 

1906  Schmidt-Nielsen,  Sigval:  Zur  Kenntnis  des  Kaseins  und  der  Labgerin- 

nung,  Upsala  Lakaref  Forh.  (N.  F.)  11,  Suppl.  Hammarsten  Fest- 
schrift No.  XV,  1-26  (1906);  through  Jahresb.  ii.  d.  Fortschr.  d  .Thier- 
chem.  36  (1906),  255,  256. 

1900  Schmiedeberg,  O. :  Ueber  die  Nucle'insaure  aus  der  Lachsmilch,  Arch.  exp. 

Path.  u.  Pharm.  43,  57-83. 

1907  Schmiedeberg,  O.:  Beitrage  zur  Kenntniss  der  tierischen  Nuclei'nsaure, 

Ibid.  57,  309-337. 

1904  Schmoll,  E.:  The  Chemical  Origin  of  Leucocytes,  Johns  Hopkins  Hosp. 

Bui.  15,  238-247. 

1875  Schmuziger,  F.:  Zur  Urinuntersuchung  bei  puerperaler  Osteomalacie, 

Centralbl.  med.  Wissensch.,  946-949. 
1895     Scholz,  Wilhelm :  Ueber  den  Einfluss  der  Schilddriisenbehandlung  auf  den 
Stoffwechsel  des  Menschen,  insbesondere  bei  Morbus  Basedowii,  Cen- 
tralbl. innere  Med.  16,  1041-1058;  1069-1083. 

1905  Scholz,  Wilhelm :  Ueber  den  Stoffwechsel  der  Cretinen,  Zeit.  exp.  Path.  u. 

Ther.  2,  271-384. 
(1911)  Schottin,  W.  R.:  Phosphorarmut  der  Rindenzentren  Ursache  der  Migrane, 

Heilung  durch  Phosphorlecithin-Oel,  Med.  Klinik   (Dresden)    9,  339- 

341;  through  Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  41  (1911),  486. 
1893-4  Schoumow-Simanowsky,  E.  O. :  Ueber  den  Magensaf t  und  das  Pepsin 

bei  Hunden,  Arch.  exp.  Path.  u.  Pharm.  33,  336-352. 

1901  Schreiber,  E.:  Ueber  die  Verwendung    des    frischen    Kaseins    in    der 

Ernahrung,  Centralbl.  Stoffwechsel-  u.  Verdauungskrankh.  2,  115-118. 
1897     Schreiber  and  Waldvogel:  Ueber  Sanose,    ein    neues    Eiweisspraparat, 
Deut.  med.  Wochenschr.  23,  Therapeut.  Beilage,  65-67. 

1876  Schrodt,   Max:   Vergleichende   Knochenuntersuchungen,    angestellt    am 

Skelete  eines  Fleischfressers,  Landwirtsch.  Versuch.  Stat.  19,  349-400. 
1889     Schrodt,  Max:  Ueber  die  Zusammensetzung  von  Buffelmilch,  Vierteljahr- 

esschr.  ii.  d.  Fortschr.  d.  Chem.  d.  Nahrungs-  u.  Genussmittel  4,  137, 

138. 
1885     Schrodt,  M.,  and  H.  Hansen:  Ueber  die  Zusammensetzung  der  Aschen 

von  Kuhmilch,  Landwirtsch.  Versuch.  Stat.  31,  55-83. 

1905  Schroder,  G.:  Ueber  neuere  Medikamente  und  Nahrmittel  fur  die  Be- 

handlung  der  chronischen  Lungentuberkulose,  Zeit.  Tuberkulose  u. 
Heilstattewesen  7,  238-242. 

1906  Schroder,  G.:  Ueber  neuere  Medikamente  und  Nahrmittel  fur  die  Be- 

handlung  der  chronischen  Lungentuberkulose,  Ibid.  8,  509-518. 

1887  Schroter,  R. :  Untersuchungen  iiber  die  Eiweisskorper  der  Menschen-  und 
Kuhmilch,  Jahrb.  Kinderheilk.  n.  s.  26,  362-371. 

1913  Schryver,  S.  B.:  Some  Investigations  on  the  Phenomena  of  "Clot"  Form- 
ation. Pt.  I.  On  the  Clotting  of  Milk,  Proc.  Roy.  Soc.  London  (B)  86, 
460-481. 


686  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1890  Schtscherbak,  A.:  Beitrage  zur  Lehre  von  der  Abhangigkeit  des  Phos- 
phorumsatzes  von  gesteigerter  oder  herabgesetzter  Gehirnthatigkeit, 
Diss.,  St.  Petersburg;  through  Neurolog.  Centralbl.  10  (1891),  171, 172. 

1878  Schiitzenberger,  P.:  Faits  pour  servir  a  l'histoire  de  la  levure  de  biere, 
Compt.  rend.  Acad,  des  sci.  78,  493. 

1903  Schulte-Bauminghaus,  Clemens:  Ueber  die  Wirkung  und  den  Verbleib 

einiger  an  Milchkuhe  gefutterten  Mineralstoflverbindungen.  (Eisen, 
Calcium,  Chlor,  Phosphorsaure.)  Mitteil.  d.  landwirtsch.  Inst.  d. 
kbnig.  Univ.  Breslau  2,  25-69. 

1899  Schulz,  Fr.  N.,  and  O.  Falk :  Phosphorsaureausscheidung  nach  Castration, 
Zeit.  physiol.  Chem.  27,  250-254. 

1901  Schulz,  Fr.  N.,  and  J.  Mainzer:  Ueber  den  Verlauf  der  Phosphorsaure- 
ausscheidung beim  Hunger,  Ibid.  32,  268-277. 

1895  Schulze,  E.:  Ueber  die  Bestimmung  des  Lecithingehaltes  der  Pflanzen- 

samen,  Ibid.  20,  225-232. 
1897     Schulze,  E.:    Ueber  den    Lecithingehalt    einiger    Pflanzensamen    und 

Oelkuchen,  Landwirtsch.  Versuch.  Stat.  49,  203-214. 
1907     Schulze,  E.:  Ueber  den  Phosphorgehalt  einiger  aus  Pflanzensamen  dar- 

gestellter  Lecithinpraparate,  Zeit.  physiol.  Chem.  52,  54-61. 
1908a  Schulze,  E. :  Ueber  die  Darstellung  von  Lecithin  und  anderen  Phosphatid- 

en  aus  Pflanzensamen  verwendbaren  Methoden,  Ibid.  55,  338-351. 
1908b  Schulze,  E.:  Ueber  pflanzliche  Phosphatide,  Chem.-Zeitung  32,  981-983. 

1904  Schulze,  E.,  and  N.  Castoro:  Findet  man  in  Pflanzensamen  und  in  Keim- 

pflanzen  anorganische  Phosphate?  Zeit.  physiol.  Chem.  41,  477-484. 

1894  Schulze,  E.,  and  S.  Frankfurt :  Ueber  den  Lecithingehalt  einiger  vegetab- 

ilischer  Substanzen,  Landwirtsch.  Versuch.  Stat.  43,  307-318. 

1891a  Schulze,  E.,  and  A.  Likiernik:  Darstellung  von  Lecithin  aus  Pflanzen- 
samen, Ber.  deut.  chem.  Gesell.  24,  71-74. 

1891b  Schulze,  E.,  and  A.  Likiernik:  Ueber  das  Lecithin  der  Pflanzensamen, 
Zeit.  physiol.  Chem.  15,  405-414. 

1911  Schulze,  E.,  and  U.  Pfenninger:  Untersuchungen  iiber  die  in  den  Pflanzen 
vorkommenden  Betaine.      I.  Mitteilung,  Ibid.  71,  174-185. 

1889  Schulze,  E.,  and  E.  Steiger:  Ueber  den  Lecithingehalt  der  Pflanzensamen, 
Ibid.  13,  365-384. 

1896  Schulze,  E.,  and  E.  Winterstein:  Ueber  einen  phosphorhaltigen  Bestand- 

theil  der  Pflanzensamen,  Ibid.  22,  90-94. 

1903  Schulze,  E.,  and  E.  Winterstein:  Beitrage  zur  Kenntnis  der  aus  Pflanzen 

darstellbaren  Lecithine  (Erste  Mittheilung.)  Ibid.  40,  101-119. 
1906     Schumoff-Simanowski,  C,  and  N.  Sieber:  Das  Verhalten  des  Lecithins 
zu  fettspaltenden  Fermenten,  Ibid.  49,  50-63. 

1895  Schwartz,  Emil :  Ueber  die  Phosphorsaurestoffwechsel  bei  der  Pneumonle, 

Wien.  med.  Blatter  18,  775-778;  792-794;  807,  808. 
1894     Schwiening,  Heinrich:  Ueber  fermentative  Prozesse    in    den    Organen, 
Arch.  path.  Anat.  u.  Physiol.  136,  444-481. 

1904  Scofone,  L.:  Sul  comportamento  della  fitina  nelP  organismo,  Giornale  d. 

reale  Accad.  di  med.  di  Torino  10  (ser.  4),  630,  631. 
1899     Scott,  F.  H.:  On  the  Structure,  Microchemistry    and     Development     of 

Nerve  Cells,  with  Special  Reference  to     their     Nuclein     Compounds, 

Trans,  of  the  Canadian  Inst.  6,  405-438.  88  refs. 
1906    Scott,  F.  H. :  On  the  Liberation  of  Phosphorus  Compounds  from  Nucleins, 

British  Med.  Jour.,  1791,  1792. 


PHOSPHORUS  METABOLISM  687 

1889  Sebelien,  John:  (Ueber  Peptone  und  ahnliche  Substanzen),  Tidsskrift  for 
Physik  og  Chemi  9  (ser.  2),  234-256;  through  Biedermann's  Centralbl. 
f.  Agrictilturchem.  18  (1889),  717,  718. 

1894  Sebelien,  John:  Ueber  das  bei  der  Pepsindigestion  des  Caseins  abge- 

spaltene  Nuclei'n,  Chemiker  Zeitung  18,  2018. 

1895  Sebelien,  John:  Ueber  das  Verhalten  des  bei  der  Pepsindigestion  des 

Caseins  abgespaltenen  Pseudonucleins,  Zeit.  physiol.  Chem.  20,  443-454. 
1904     Secheret,  Georges:  Contribution  a.  l'etude  de  la  medication  phosphoree. 
Etude  therapeutique  et  clinique  de  l'acide  anhydro-oxy-methylene-  di- 
phosphorique,  These  de  Paris.       131  pp. 

1899  Sedlmair,  August  Carl:  Ueber  die  Abnahme  der  Organe,  insbesondere 

der  Knochen,  beim  Hunger,  Zeit.  Biol.  37,  (n.  s.  19),  25-58. 

1900  von  Seelhorst,  C,  N.  Georgs  and  F.  Fahrenholtz:  Einfluss  des  Wassers- 

gehaltes  und  der  Dungung  des  Bodens   auf  die  Production  und  die 
Zusammensetzung  von  Futterpflanzen     italienisches     Raigrass     und 
Klei,  Jour.  f.  Landwirtsch.  48,  265-286. 
1879    Seeman:  Zur  Pathogenese  und  Aetiologie  der  Rachitis,  Arch.  path.  Anat. 
u.  Physiol.  77,  299-315. 

1897  Senator,  H.:  Zur  Kenntniss  der  Osteomalacic  und  der  Organotherapie, 

Berlin,  klin.  Wochenschr.  34,  109-112;  143,  144. 
1888    Sendtner,  J.:  Zur  Phosphaturie,  Munch,  med.  Wochenschr.,  671,  672. 
1897a  Serono,  Cesare:  Sur  les  injections  de  lecithine  chez  l'homme  et  chez  les 

animaux.      Note  preventive.      Arch.  ital.  de  biol.  27,  349-354. 
1897b  Serono,  Cesare:  Ricerche  sulle  iniezioni  di  lecitina,  La  Riforma  medica, 

Napoli,  3,  616-618;  627-630;  637-639. 
1902    Serono,  Cesare:  Considerazioni  terapeutiche  sull'  uso  della  lecitina,  Gaz- 

zetta  degli  ospedali  (Milan)  23,  II,  1246-1248. 
1911     Serono,  Cesare,  and  Antonietta  Palozzi:  Sui  lipoidi  contenuti  nel  tuorlo 

d'uovo,  Archivio  di  farmacol.  speriment.  11,  553-570;  through  Chem. 

Abs.  5  (1911),  2858. 
1913    Serono,  Cesare,  and  Antonietta  Palozzi:  Sui  lipoidi  contenuti  nella  sos- 

tanza  nervosa,  Ibid.  15,  375-395, 

1898  Setti,  Giovanni:  Ricerche  urologiche  in  due  casi  di  bronco-pneumonite 

acuta,  II  Policlinico  5,  425-436. 

1911  Schackell,  L.  F.:  Phosphorus  Metabolism  during  Early  Cleavage  of  the 
Echinoderm  Egg,  Science  34,  573-576. 

1902  Sherman,  Henry  C:  Experiments  on  the  Metabolism  of  Nitrogen,  Sul- 
phur and  Phosphorus  in  the  Human  Organism,  U.  S.  Dept.  Agr.,  Office 
Exp.  Sta.  Bui.  121,  47  pp. 

1908  Sherman,  Henry  C. :  Experiments  upon  the  Metabolism  of  Phosphorus  in 
Man,  Proc.  Amer.  Soc.  Biol.  Chem.  1,  135,  136. 

1911     Sherman,  Henry  C. :  Chemistry  of  Food  and  Nutrition,  New  York.  355  pp. 

1900  Sherman,  H.  C,  and  P.  B.  Hawk:  On  the  Elimination  of  Nitrogen,  Sul- 
phates and  Phosphates  after  the  Ingestion  of  Proteid  Food,  Amer. 
Jour.  Physiol.  4,  25-49. 

1910  Sherman,  H.  C,  Arthur  J.  Mettler,  and  J.  Edwin  Sinclair:  Calcium,  Mag- 
nesium and  Phosphorus  in  Food  and  Nutrition,  U.  S.  Dept.  Agr.,  Office 
Exp.  Sta.  Bui.  227.      70  pp. 

1910  Shibayama,  Gorosaku:  Some  Observations  Concerning  Beriberi,  Philip- 
pine Jour,  of  Sci.,  B,  5,  123-125. 


688  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1857     Sick,  Paul:  Versuche  iiber  die  Abhangigkeit  des  Phosphorsauregehaltes 

des  Urins  von  der    Phosphorsaurezufuhr,    Arch,     physiol.     Heilk.     1 

(n.  s.),  482-503. 
1909a  Sieber,  Nadina:  Zur  Frage  der  Einwirkung  von  Alkohol  auf  den  Gehalt 

der  tierischen  Organe  an  Phosphatiden,  Biochem.  Zeit.  23,  304-323. 
1909b  Sieber,  Nadina:  Die  Lipoide  der  Lunge,  Zeit.  physiol.  Chem.  62,  250-253. 
1909     Sieber,  N.,  and  W.  Dzierzgowski :  Die  Zusammensetzung  der  Lunge,  Ibid. 

62,  254-258. 
1879     Siedamgrotsky   and   Hofmeister:   Die   Einwirkung   andauernder   Milch- 

saureverabreichung  auf  die  Knochen  der  Pflanzenfresser,  Arch,  wis- 

sensch.  u.  prakt.  Thierheilk.  5,  243-282. 

1898  Siegert,  F.:  Ueber  typische  Osteomalacie  im  Kindesalter,  Munch,  med. 

Wochenschr.  45,  1401-1404. 
3893     Siegfried,  Max:  Ueber  eine  neue  stickstoffhaltige  Saure  der  Muskeln, 
Ber.  d.  k.  Sachs.  Gesell.  d.  Wissensch.,  (1893),  484-489. 

1894  Siegfried,  Max:  Ueber  Fleischsaure,  Arch.  Anat.  u.  Physiol.,  physiol. 

Abt.,  401-418. 

1895  Siegfried,  Max:  Ueber  Phosphorfleischsaure,  Ber.  deut.  chem.  Gesell.  28r 

515-519. 

1896  Siegfried,  Max:  Zur  Kenntniss  der  Phosphorfleischsaure,  Zeit.  physiol. 

Chem.  21,  360-379. 

1897  Siegfried,  Max:  Zur  Kenntniss  des  Phosphors  in  der  Frauen-  und  Kuh- 

milch,  Ibid.  22,  575-578. 

1899  Siegfried,  Max:  Zur  Kenntniss  der  Extractivstoffe  des  Muskels,  Ibid.  28, 

524-529. 

1905  Siegfried,  M.,  and  H.  Mark:  Zur  Kenntniss  des  Jecorins,  Ibid.  46,  492- 

496. 
1904     Siegmann,  J.:  Ueber  die  Anwendung  des  Protylin  "Roche"  bei  Neurosen, 

Aerztlichen  Central-Zeitung,  therapeut.  Ratgeber  16,  9-11. 
1868     Siewert,  M.:  Ueber  den  Stickstoffumsatz  der  im  Korper  verbrauchten 

Eiweisskorper,  Zeit.  ges.  Naturwissensch.  31,  458-480. 

1906  Sikes,  Alfred  W.:   On  the  Phosphorus  and   Calcium   of  Human   Milk, 

Thesis,  London;  also  Jour,  of  Physiol.  34,  464-480. 
1904     Silberstein,  James:  Ueber  die  Anwendung  organischer  Phosphorprapar- 
ate  in  Form  des  Hamoprotagons,  Deut.  Med.-  Zeitung,  289-  291. 

1902  Simnitski,  S.,  and  P.  Rodoslawow:  (Beitrag  zur  Urologie  des  Ikterus), 

Wratschetnaja  gazetta  1902,  No.  40-42;  through  Jahresb.  u.  d. 
Fortschr.  d.  Thierchem.  32  (1902),  672. 

1911  Simon,  Friedrich:  Zur  Kenntnis  der  Autolyse  des  Gehirns,  Zeit.  physiol. 
Chem.  72,  463-483. 

1911  Simpson,  G.  C.  E.,  and  E.  S.  Edie:  On  the  Relation  of  the  Organic  Phos- 
phorus Content  of  "Various  Diets  to  Diseases  of  Nutrition,  Particular- 
ly Beriberi.  I.,  Annals  of  Trop.  Med.  and  Parasitology  5,  313-345; 
through  Chem.  Abs.  6  (1912),  1458. 

1901  Siven,  V.  A.:  Zur  Kenntniss  des  Stoffwechsels  beim  erwachsenen  Mensch- 
en,  mit  besonderer  Beriicksichtigung  des  Eiweissbedarfs,  Skand. 
Arch.  Physiol.  11,  308-332. 

1903  Siwertzow,   D.   I.:    (Vergleichende  Untersuchungen   iiber   den  Lecithin- 

gehalt  bei  menschlichen  Embryonen  und  bei  Kindern  im  fruhen 
Lebensalter),  Diss.,  St.  Petersburg;  through  Biochem.  Centralbl.  2 
(1904),  310,  311. 


PHOSPHORUS  METABOLISM  689 

1908  Sleeswyk,  J.  G.:  Ueber  die  angebliche  bakteriolytische  Eigenschaft  des 

Lecithins  und  iiber  die  Immunisierungs  mittels  Lecithin-Typhustoxine, 
Deut.  med.  Wochenschr.  34,  II,  2263-2265. 

1906a  Slowtzoff,  B.:  Ueber  die  Resorption  des  Lecithins  aus  den  Darmkanal, 
Beitrage  z.  chem.  Physiol,  u.  Path.  7,  509-513. 

Other  spellings  of  this  author's  name  are:  Slowtzow,  Slowzow,  and 
Slovcov. 

1906b  Slowtzoff,  B.:  Die  Wirkung  des  Lecithins  auf  den  Stoffwechsel,  Ibid. 
8,  370-388. 

1909a  Slowtzoff,  B.:  (Ueber  den  Einfluss  der  Fischnahrung  auf  den  Phosphor-, 
Kalk-  und  Magnesiaumsatz),  Verhandl.  Gesell.  Russ.  Aerzte,  St.  Peters- 
burg 76,  220;  through  Zentralbl.  Biochem.  u.  Biophys.  10  (1910),  375, 
376. 

1909b  Slowtzoff,  B.:  (Der  Phosphor-,  Calcium-  und  Magnesia- Stoffwechsel  bei 
Ersatz  der  Fleischnahrung  durch  Fisch),  Russki  Wratsch,  8,  1232- 
1234;  through  Jahresb.  u.  d.  Fortschr.  d.  Thierchem.  39   (1909),  606. 

1911  Smetanka,  Franz:  Zur  Herkunft  der  Harnsaure  beim  Menschen.      Ein 

Beitrag  zur  Physiologie  der  Drusen,  Arch.  ges.  Physiol.  138,  217-274. 
1889     Smith,  Fred:  The  Chemistry  of  the  Urine  of  the  Horse,  Proc.  of  the 
Royal  Soc.  46,  328-340. 

1912  Smith,  J.  Lorrain,  and  W.  Mair:  The  Development  of  Lipoids  in  the  Brain 

of  the  Puppy,  Jour,  of  Path,  and  Bacteriol.  17,  123-126. 

1904  Smolenski,  J.:  O  znaczeniu  leczniczen  zwaizkow  glicerynofosforowych  u 

dzieci,  (Ueber  der  therapeutischen  Wert  der  Glycerophosphatverbind- 
ungen  bei  Kindern),  Medycyna,  Warszawa,  32,  733;  756;  777;  through 
Jahrb.  Kinderheilk.  61  (1904),  680. 

1909  Smolenski,  K.:  Zur  Kenntnis  der  aus  Weizenkeimen  darstellbaren  Phos- 

phatide. V.  Mitteilung.  Ueber  Phosphatide.  (Winterstein's  series.) 
Zeit.  physiol.  Chem.  58,  522-526. 

1905  Snowman,  J. :  The  Therapeutic  Action  of  Glycerophosphates  in  Combina- 

tion with  Albumens,  Birmingham  Med.  Review  58,  519-526. 
1888     Soldner,  F.:  Die  Salze  der  Milch  und  ihre  Beziehungen  zu  dem  Verhalten 

des  Caseins,  Landwirtsch.  Versuch.  Stat.  35,  351-436. 
1902    Soldner,  F.:  Die  Aschenbestandtheile  des  neugeborenen  Menschen  und 

der  Frauenmilch,  Verhandl.  d.  Gesell.  f.  Kinderheilk.  19,  154-160. 
1902     Soetbeer,  Franz:  Ueber  Phosphaturie,  Jahrb.  Kinderheilk.  56,  1-10. 
1902     Soetbeer,  Franz,  and  Hans  Krieger:  Ueber  Phosphaturie,  Deut.  Arch. 

klin.  Med.  72,  553-559. 
1912     Sokolov,  S.  K.:  (Assimilation  of  Sodium  Nucleinate  in  Old  and  Young 

Organisms),  Russki  Wratch   11,  1065-1067;   through   Chem.   Abs.   7 

(1913),  1896. 
1909    Soli,  Ugo:  Modifications  du  developpement  des  os    chez    les    animaux 

prives  de  thymus,  Archives  ital.  de  biol.  52,  217-224;  through  Chem. 

Abs.  5  (1911),  2128. 
1902    Somerfeld,  Paul,   and  Wilhelm  Caro:  Zur  Kenntniss  der  Ausniitzung 

von  Phosphor  und  Stickstoff  bei  reiner  Milchernahrung  alterer  Kinder, 

Arch.  Kinderheilk.  33,  161-166. 
1880    Sotnitschewsky:  Glycerinphosphorsaure  im  normal  en  menschlichen  Harn, 

Zeit.  physiol.  Chem.  4,  214-216. 


690  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1878  Soxhlet,  F.:  Untersuchungen  iiber  den  Stoffwechsel  des  Saugkalbes, 
Erster  Ber.  u.  d.  Arbeiten  d.  K.  K.  landwirtsch.  Versuchsstat.  z.  Wien. 
1878,  101-105;  also  Oesterreich.  landwirtsch.  Wochenbl.  4  (1878),  290- 
292,  315,  316,  327,  328;  through  Jahresb.  ii.  d.  Fortschr.  d.  Thierchem. 
8  (1878),  333-337  and  Centralbl.  f.  agr.  Chem.  7  (1878),  748-752,  887- 
891. 

1893  Soxhlet,  F.:  Die  chemischen  Unterschiede  zwischen  Kuh-  und  Frauen- 

milch  und  die  Mittel  zu  ihrer  Ausgleichung,  Munch,  med.  Wochenschr. 
40,  61-65. 
1903     Soxhlet,  F.:  Bezeichnung  und  Beschaffenheit  des  fur  Futterungszwecke 
dienenden  phosphorsauren  Kalks,     Landwirtsch.    Versuch.     Stat.     58, 
376,  377. 

1882  Speck,  Karl:  Untersuchungen  iiber  die  Beziehungen  der  geistigen  Thatig- 

keit  zum  Stoffwechsel,  Arch.  exp.  Path.  u.  Pharm.  15,  81-145. 
1906     Spiro,  K.:  Beeinflussung  und  Natur  des  Labungsvorganges,  Beitrage  z. 

chem.  Physiol,  u.  Path.  8,  365-369. 
1899     Spitzer,  W.:  Die  Ueberfuhrung  von  Nucleinbasen  in  Harnsaure  durch 

die  sauerstoffubertragende  Wirkung  von  Gewebsausziigen,  Arch.  ges. 

Physiol.  76,  192-203. 

1894  Springer,  Maurice:  Etude  physiologique  et  chimique  sur  la  croissance, 

Semaine  med.  14,  390-393. 
1902     Springer,  Maurice:  L'energie  de  croissance  et  les  lecithines     dans     les 

decoctions  de  cereales,  Paris.      170  pp. 
1866     Ssubotin:  Ueber  den  Einfluss  der  Nahrung  auf  die  quantitative  Zusam- 

mensetzung  der  Milch,  Arch.  path.  Anat.  u,  Physiol.  36,  561-570. 

1883  Stadelman,  E.:  Ueber  die  Ursachen  der  pathologischen  Ammoniakaus- 

scheidung  beim   Diabetes   mellitus   und   des   Coma   diabeticum,   Exp. 

Arch.  17,  419. 
1906     Stanek,  Vladimir:  Ueber  die  quantitative  Bestimmung  von  Cholin  und 

Betain  in  pflanzlichen  Stoffen  und  einige  Bemerkungen  iiber  Lecithine, 

Zeit.  physiol.  Chem.  48,  334-346. 
1845     Stark,  James:  On  the  Bones.  Part    I.  Chemical  Constitution  of  the  Bones 
of  the  Vertebrated  Animals,  Edinburgh  Med.  and  Surg.  Jour.  63,  308- 

325. 

1910  Starkenstein,  Emil:  Die  biologische  Bedeutung  der  Inositphosphorsaure, 

Biochem.  Zeit.  30,  56-98. 

1911  Starkenstein,  Emil:  Ionenwirkung  der  Phosphorsauren,  Ibid.  32,  243-265. 
1902a  Stassano,  H.,  and  F.  Billon:  Contribution  a  la  connaissance  de  Taction  de 

la  lecithine  sur  les  leucocytes,  Compt.  rend.  Soc.  de  biol.  54,  167-169. 
1902b  Stassano,  H.,  and  F.  Billon :  Sur  la  leucocytose  produite  dans  le  peritoine 

par  les  injections  de  lecithine,  Ibid.  54,  169,  170. 
1902c  Stassano,  H.,  and  F.  Billon:  Contribution  a  la  connaissance  de  Taction  de 

la  lecithine  sur  les  elements  figures  du  sang,  Compt.  rend.  Acad,  des 

sci.  134,  318-321. 
1902d  Stassano,  H.,  and  F.  Billon:  Sur  la  diapedese  des  leucocytes  charges  de 

lecithine  et  sur  Tabsorption  de  la  lecithine  par  l'endothelium  vasculaire, 

Ibid.  134,  430-432. 
1903a  Stassano,  H.,  and  F.  Billon:  La  lecithine  n'est  pas  dedoublee  par  le  sue 

pancreatique  meme  kinase,  Compt.  rend.  Soc.  de  biol.  55,  482,  483. 
1903b  Stassano,  H.,  and  F.  Billon:  La  lecithine  pure     ingeree     se     retrouve 

inalteree  dans  la  lymphe  provenant  des  chyliferes,  Ibid.  55,  924-926. 


PHOSPHORUS  METABOLISM  691 

1907a  Steel,  Matthew,  and  William  J.  Gies:  On  the  Use  of  Bone  Ash  with  the 

Diet,  in  Metabolism  Experiments  on  Dogs,  Amer.  Jour.  Physiol.  20, 

343-357. 
1907b  Steel,  Matthew,  and  William  J.  Gies:  On  the  Chemical  Nature  of  Para- 

nucleoprotagon,  a  New  Product  from  Brain,  Ibid.  20,  378-398. 
1869     Steiner,  Jacob:  Ueber  die  pathologisch-anatomischen  Veranderungen  bei 

der  Osteomalacie,  Inaug.  Diss.,  Zurich.      50  pp. 
1898     Steinitz,  Franz :  Ueber  das  Verhalten  phosphorhaltiger  Eiweisskorper 

im  Stoffwechsel,  Arch.  ges.  Physiol.  72,  75-104. 
1900     Steinitz,  Franz:  Ueber  Versuche  mit    kiinstlicher     Ernahrung,     Inaug. 

Diss.,  Breslau.      44  pp. 

1903  Steinitz,  Franz:  Zur  Kenntnis  der  chronischen  Ernahrungsstorungen  der 

Sauglinge,  Jahrb.  Kinderheilk.  57,  689-730. 

1904  Steinitz,  Franz :  Ueber  den  Einfluss  von  Ernahrungsstorungen  auf  die 

chemische  Zusammensetzung  des  Sauglingskorpers,  Ibid.  59,  447-461. 

1904  Steinitz,  Franz,  and  Richard  Weigert :  Demineralisation  und  Tuberkulose, 

Vorlaufige  Mitt.  Deut.  med.  Wochenschr.  30,  I,  838,  839. 

1905  Steinitz,  Franz,  and  Richard     Weigert:     Ueber     Demineralisation    und 

Fleischtherapie  bei  Tuberkulose,  Jahrb.  Kinderheilk.  61,  147-161. 
1900  a  von  Stejskal,  C,  and  F.  Erben:  Klinisch-chemische  Studien:  Stoffwech- 

selversuch  bei  lymphatischer  und  lional-myelogener  Leukamie,  Zeit. 

klin.  Med.  39,  151-170. 
1900b  von  Stejskal,  C,  and  F.  Erben:  Klinisch-chemische  Studien:  II.  Stoff- 

wechselversuch  bei  pernicioser  Anamie,  Ibid.  40,  165-180. 
1897     de  Stella,  H.:  Les  glycerophosphates.       Leur  influence  sur  la  nutrition 

intime  et  leur  role  physiologique  dans  l'organisme,  Archives  internat. 

de  pharmacodyn.  et  de  ther.  3,  351-357. 
1890     Stellwag:  August:  Die  Zusammensetzung  der  Futtermittelfette,  Land- 

wirtsch.  Versuch.  Stat.  37,  135-154. 
(1913)  Stephan,  Alfred:  (The  Physiology  of  Nuclein  Metabolism),  Apoth.  Zei- 

tung  27,  816-819. 
1909     Stepp,   Wilhelm:   Versuche   uber  Futterung   mit   lipoidfreier   Nahrung, 

Biochem.  Zeit.  22,  452-460. 
1911     Stepp,  Wilhelm :  Experimentelle  Untersuchungen  liber  die  Bedeutung  der 

Lipoide  fur  die  Ernahrung,  Zeit.  Biol.  57  (N.  F.  39),  135-170. 
1913a  Stepp,  .Wilhelm :  Weitere  Untersuchungen  liber  die  Unentbehrlichkeit  der 

Lipoide  fiir  das  Leben.       Ueber  die  Hitzezerstorbarkeit  lebenswicht- 

iger  Lipoide  der  Nahrung,  Ibid.  59  [N.  F.  41],  366-395. 
1913b  Stepp,  Wilhelm:  Fortgesetzte  Untersuchungen  liber  die  Unentbehrlich- 
keit der  Lipoide  fiir  das  Leben.      Ueber  das  Verhalten  der  lebenswich- 

tigen  Stoffe  zu  den  Lipoidextraktionsmitteln,  Ibid.  62  [N.  F.  44],  405- 

417. 
1907     Stern,  M.,  and  H.  Thierf elder:  Ueber  die  Phosphatide  des  Eigelbs,  Zeit. 

physiol.  Chem.  53,  370-385. 
1893     Sternberg,  Maximilian:  Ueber  Diagnose  und  Therapie  der  Osteoinalacie, 

Zeit.  klin.  Med.  22,  265-313. 
1904     Steudel,  H.:  Zur  Kenntnis  der  Thymusnucleinsauren.       I.   Mitt.,   Zeit. 

physiol.  Chem.  42,  165-170. 
1905a  Steudel,  H. :  Zur  Kenntnis  der  Thymusnucleinsauren.      II.  Mitt.,  Ibid.  43, 

402-405. 


692  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1905b  Steudel,  H.:  Zur  Kenntnis  der  Thymusnucleinsauren.       III.  Mitt.,  Ibid. 

46,  332-336. 
1906a  Steudel,  H. :  Ueber  die  Oxydation  der  Nucleinsaure.      I.  Mitteilung,  Ibid. 

48,  425-429. 
1906b  Steudel,  H.:  Die  Zusammensetzung  der  Nucleinsauren  aus  Thymus  und 

aus  Heringsmilch,  Ibid.  49,  406-409. 
1907a  Steudel,  H.:  Nucleine,  Nucleinsauren  und  ihre  Spaltungsprodukte,  Bio- 

chem.  Centralbl.  6,  125-134. 
1907b  Steudel,  H.:  Ueber  die  Oxydation  der  Nucleinsaure.  II.  Mitteilung,  Zeit. 

physiol.  Chem.  50,  538,  539. 
1907c  Steudel,  H.:  Zur  Analyse  der  Nucleinsauren,  Ibid.  52,  62. 
1907d  Steudel,  H.:  Die  Zusammensetzung  der  Nucleinsauren  aus  Thymus  und 

aus  Heringssperma,  Ibid.  53, 14-18. 
1907e  Steudel,  H. :  Ueber  die  Bildung  von  Pyrimidinderivaten  aus  Purinkorpern, 

Ibid.  53,  508-513. 
1907f    Steudel,  H.:  Ueber  die  Guanylsaure  aus  der  Pankreasdriise,  Ibid.  53,. 

539-544. 
1908a  Steudel,  H.:  Ueber  die  Kohlenhydratgruppe  in    der    Nucleinsaure.       I. 

Mitt,  Ibid.  55,  407-411. 
1908b  Steudel,  H.:  Ueber  die  Kohlenhydratgruppe  in  der  Nucleinsaure.       IL 

Mitt.,  Ibid.  56,  212-219. 
1911a  Steudel,  H.:  Zur  Histochemie  der  Spermatozoen,  Ibid.  72,  305-312. 
1911b  Steudel,  H.:  Zur  Histochemie  der  Spermatozoen.       II.  Mitteilung,  Ibid. 

73,  471-477. 
1912     Steudel,  H.:  Ueber  den  Bau  der  Nucleinsaure  aus  der  Thymusdriise,  Ibid. 

77,  497-507. 
1913a  Steudel,  H.:  Zur  Histochemie  der  Spermatozoen.      II.  Mitteilung,  Ibid. 

83,  72-78. 
1913b  Steudel,  H.:  Ueber  das  Nucleohiston.      I.  Mitteilung,  Ibid.  87,  207-213. 

1910  Steudel,  H.,  and  P.  Brigl.:  Ueber  die  Guanylsaure  aus  der  Pankreasdriise* 

II.  Mitt.,  Ibid.  68,  40-51. 

1911  Steudel,  H.,  and  P.  Brigl:  Ueber  die  Thyminsaure,  Ibid.  70,  398-403. 
1889     Stilling,  H.,  and  J.  v.  Mering:  Ueber  experimented  Erzeugung  der  Os- 
teomalacic, Centralbl.  med.  Wissensch.  27,  803,  804. 

1908  Stoelzner,  Helene:  Ueber  den  Einfluss  von  Strontiumverfutterung  auf 
die  chemische  Zusammensetzung  des  wachsenden  Knochens,  Biochem. 
Zeit.  12,  119-137. 

1903  Stoelzner,  Wilhelm:  Die  Einwirkung  das  Phosphors  auf  den  rachitischen 

Knochenprozess,  Verhandl.  d.  Gesell.  deut.  Naturforsch.  u.  Aerzte  z. 
Kassel,  1903,  264-266;  also  Verhandl.  d.  Gesell.  f.  Kinderheilk.,  Wies- 
baden, 20  (1903),  198-205. 

1904  Stoelzner,  Wilhelm:  Pathologie  und  Therapie  der  Rachitis,  Berlin.  176 

pp.       20%  pp.  refs. 
1908     Stoelzner,  Wilhelm:  Die  zweifache  Bedeutung  des  Calciums     fiir     das 

Knochenwachstum,  Arch.  ges.  Physiol.  122,  599-604. 
1869     Stohmann,    F.:    Ueber   Knochenbriichigkeit    erzeugendes    Heu,    Zeit.    d. 

landwirtsch.  Central- Vereins  d.     Prov.     Sachsen,     1869,     9;     through 

Jahresb.  u.  d.  Fortschr.  d.  Agriculturchem.,  1868-9,  546,  547. 
1895     Stoklasa,  Julius:  Chemische  Untersuchungen  auf  dem  Gebiete  der  Phyto- 

pathologie,  Zeit.  physiol.  Chem.  21,  79-86. 


PHOSPHORUS  METABOLISM  693 

1896a  Stoklasa,  Julius:  Ueber  die  physiologische  Bedeutung  des  Lecithins  in 

der  Pflanze,  Ber.  deut.  chem.  Gesell.  29,  2761-2771. 
1896b  Stoklasa,  Julius:  Ueber  die  Verbreitung  und  physiologische  Bedeutung 

des  Lecithins  in  der  Pflanze,  Sitzungsber.  d.  math,  -naturw.  Classe  d. 

kais.  Akad.  d.  Wissensch.  105,  Abt.  1,  604-632. 
1897     Stoklasa,  Julius:  Zur  Kenntniss  des  Phosphors  in  der  Frauen-  und  Kuh- 

milch,  Zeit.  physiol.  Chem.  23,  343-346. 

1876  Stokvis,  Barend  Josephus:  (Weitere  Beitrage  zur  Kenntniss  der  Phos- 

phorsaureausscheidung  bei  Arthritis),     Weekblad     van     het     Nederl. 

Tijdschr.  f.  Geneesk.,  1876,  No.  36,  565;  through  Jahresb.  ii.  d.  Fortschr. 

d.  Thierchem.  6  (1876),  156. 
1897     Storch,  Karl:  Beitrage  zur  Kenntniss  der  Eiweisskorper  der  Kuhmilch, 

Monatsh.  f.  Chem.  18,  244-281. 
1902     Storch,  Karl:  Beitrage  zur  Kenntnis  des  Caseinogens  der  Eselinmilch, 

Ibid.  23,  712-730. 
1899     Straub,  W.:  Ueber  den  Einfluss  der  Wasserentziehung  auf  den  Stoff- 

wechsel  und  Kreislauf,  Zeit.  Biol.  38  (N.  F.  20),  537-566. 

1909  Straughn,  M.  N.,  and  Walter  Jones:  The  Nuclein  Ferments  of  Yeast, 

Jour.  Biol.  Chem.  6,  245-255. 
1896     Strauss,  J.:  Ueber  die  Einwirkung  des  kohlensauren    Kalkes     auf    den 

menschlichen  Stoffwechsel,  ein  Beitrag  zur  Therapie  der  harnsauren 

Nierenconcretionen  nebst  Bemerkungen  iiber  Alloxurkorperausschei- 

dung,  Zeit.  klin.  Med.  31,  493-519. 
1868     Strecker,  Adolph:  Ueber  das  Lecithin,  Annalen  der  Chim.  u.  Pharm. 

148,  77-90. 
1902     Street,  P.  W.:  The  Glycerophosphates;  Particularly  the  Glycerophosphate 

of  Sodium,  Medical  Record  (N.  Y.),  Oct.  11,  1902;  through  Jour.  Amer. 

Med.  Assoc.  39,  II,  (1902),  1072. 
1888     Strohmer,  F.:  Ueber  Biiffelmilch  und  Biiffelbutter,  Zeit.  Nahrungsmittel- 

unters.  u.  Hygiene  2,  17-19;  through  Jahresb.  ii.  d.  Fortsch.  d.  Thier- 
chem. 18  (1888),  104. 
1912     Strong,  R.  P.,  and  B.  C.  Crowell:  The  Etiology  of  Beriberi,  Philippine 

Jour,  of  Sci.,  B,  7,  271-413;  through  Exp.  Sta.  Record  28  (1913),  763, 

764. 

1877  Striibing:  Ueber  die  Phosphorsaure  im  Urin  unter  dem  Einflusse  excit- 

irender  und  deprimirender  Mittel,  Arch.  exp.  Path.  u.  Pharm.  6,  266- 

278. 
1913a  Stuber,  B.:  Ueber  Blutlipoide  und  Phagocytose.      I.  Biochem.  Zeit.  51, 

211-223. 
1913b  Stuber,  B.:  Ueber  Blutlipoide  und  Phagocytose,  Ibid.  53,  493-500. 
1888     Stutzer,  A. :  Ueber  phosphorsaurem  Kalk  als  Viehf utter,  Chem.  Centralbl. 

59,  (19  of  ser.  3),  639. 
1908     Stutzer,  A.:  Untersuchungen  iiber  den  Gehalt  vegetabilischer  Stoffe  an 

Stickstoff,  Phosphor  und  Schwefel  in  organischer  Bindung,  Biochem. 

Zeit.  7,  471-487. 

1910  Stutzer,  A.:  Die  Bestandteile  von  Harn  und  Kot  der  wichtigeren  land- 

wirtschaftlichen  Nutztiere,  Fuhling's  landwirtsch.  Zeitung  59,  450-452; 
through  Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  40  (1910),  661,  662. 
1905    Sugar,  Martin:  Ueber    Phosphorbehandlung     der     Otosklerose,    Arch. 
Ohrenheilk.  66,  36-49. 


694  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1903  Surmont,  H.,  and  M.  Dehon:  Bilan  du  phosphore  et  de  l'azote  dans  l'ali- 

mentation  par  le  pain  complet  et  par  le  pain  blanc,  L'echo  med.  du 
nord  7,  589-591. 

1904  Surmont,  H.,  and  M.  Dehon:  Bilan  du  phosphore  et  de  l'azote  dans  l'ali- 

mentation  mixte  avec  usage  comparatif  de  pain  blanc  et  de  pain  com- 
plet, Ibid.  8,  577-579. 
1901     Suzor,  R.:  Injections  sous-cutanees  de  jaunes  d'oeufs  crus,  Compt.  rend. 

Soc.  de  biol.  53,  378,  379. 
1912     Suzuki,  U.,  T.  Shamimura  and  S.  Odake:  Ueber  Oryzanine,  ein  Bestand- 

teile  der  Reiskleie  und  seine  physiologische  Bedeutung,  Biochem.  Zeit. 

43,  89-153;  through  Exp.  Sta.  Record  28  (1913),  168. 
1907     Suzuki,  U.,  and  K.  Yoshimura:  Ueber  die  Verbreitung  von  "Anhydro-oxy- 

methylen-diphosphorsauren  Salzen"  in  Pflanzen,  Tokyo  Imper.  Univ., 

College  of  Agr.  Bui.  7,  495-502. 
1907     Suzuki,  U.,  K.  Yoshimura,  and  M.  Takaishi:  Ueber  ein  Enzym  "Phytase" 

das  "Anhydro-oxy-methylen-diphosphorsaure"  spaltet,  Ibid.  7,  503-512. 

1912  Swanson,  C.  O.:  Acidity  of  Wheat  Flour;  its  Relation  to  Phosphorus  and 

to  Other  Constituents,  Jour.  Ind.  and  Engin.  Chem.  4,  274-278. 

1913  Swanson,  C.  O. :  Chemical  Composition  of  Wheats  Compared  with  Result- 

ant Flours,  Amer.  Miller  41,  218-222;  through  Chem.  Abs.  8  (1914), 
970. 

1907  Sweet,  J.  E.,  and  P.  A.  Levene:  Nuclein  Metabolism  in  a  Dog  with  Eck's 

Fistula,  Jour,  of  Exp.  Med.  9,  229-239. 
1904-5a  Symmers,  Douglas :  A  Contribution  to  the  Knowledge  of  the  Excretion 

of  Organic  Phosphorus  in  the  Urine  in  Certain  Pathological  Conditions, 

Jour,  of  Pathol,  and  Bacteriol.  10,  159-172. 
1904-5b  Symmers,  Douglas:  An  Additional  Note  on  the  Excretion  of  Organic 

Phosphorus  in  the  Urine,  Ibid.  10,  427-430. 
1906     Szili,  Alexander:  Experimen telle  Untersuchungen  iiber   Saureintoxika- 

tion,  Arch.  ges.  Physiol.  115,  82-105. 

1892  von   Szontagh,  Felix:  Untersuchungen  iiber  den  Nucleingehalt  in  der 

Frauen-  und  Kuhmilch,  Ungar.  Arch.  Med.  1,  192-203. 

1893  von  Szontagh,  Felix:  Nucleingehalt  der  Frauen-  und  Kuhmilch,  Centralbl. 

med.  Wissensch.,  419,  420. 

1894  von  Szontagh,  Felix:  Neuere  Beitrage  zur  Kenntniss  der  Albuminsub- 

stanzeh  der  Frauen-  und  Kuhmilch,  Ungar  Arch.  Med.  3,  367-374. 
1902     von  Szontagh,   Felix:  Beitrage  zur     kunstlichen     Sauglingsernahrung, 

Jahrb.  Kinderheilk.  56,  341-349. 
1905     von  Szontagh,  Felix:  Zur  Biochemie  der  Milch,  Ibid.  62,  715-722. 
1891     Szymkiewicz,  Franz-Stanislaus:    Ueber  den  Schwefel-    und    Phosphor- 

gehalt  der  Leberzellen  des  Rindes  in  den  verschiedenen  Lebensaltern, 

Inaug.  Diss.,  Dorpat. 
1905     von  Tabora:  Ueber  die  Phosphate  des  Mageninhalts,  Zeit.  klin.  Med.  56, 

460-474. 

1908  Taghamuro,  Pasquale:  La  tabe  dorsale  e  le  iniezloni  lecitina,  Gl'  Incur- 

abili.      Giornale  di  clinica  e  di  terapia  23,  193-20 
1910     Takemura,  M.:  Ueber  Phosphorgehalt  der  Sera  bei  Gesunden,  Syphil- 
itikern  und  Carcinomkranken,  Biochem.  Zeit.  25,  508,  509. 


PHOSPHORUS  METABOLISM  695 

1913  Takeno,  J.:  Beitrage  zur  Kenntnis  des  Stoffwechsels  besonders  der  Min- 
eralien  im  Sauglingsalter.  II.  Die  Ausscheidung  der  wichtigsten 
organischen  und  anorganischen  Nahrungsbestandteile  im  Kot  unter 
wechselnden  Ernahrungs-Bedingungen,  Jahrb.  Kinderheilk.  77,  640- 
678. 

1907  Talbot,  E.  S.:  Acid  Intoxication  or  Acidosis;  a  Factor  in  Disease,  Medical 

Record,  June  1, 1907;  through  N.  Y.  Med.  Jour.,  June  8,  1907. 
(1901)  Tangl,  Franz:  (Stoffwechselversuchungen  an     Pferden  mit  kalkarmen 

Futtermitteln),  Kiserletiigyi  Kozlemenyek  4,  197;  through  Jahresb.  ii. 

d.  Fortschr.  d.  Thierchem.  31  (1901),  797. 
1902a  Tangl,  Franz:  Zur  Kenntniss  des  P-,  Ca-  und  Mg-Umsatzes  bei  Pflanz- 

enfressern,  Arch.  ges.  Physiol.  89,  227-239. 
1902b  Tangl,  Franz:  Beitrag  zur  Kenntniss  des  anorganischen  Stoffwechsels 

beim  Pferde,  Landwirtsch.  Versuch.  Stat.  57,  367—380. 
1904    Tangl,  Franz:  Der  Stoff-  und  Energieumsatz  eines  kiinstlich  ernahrten 

Sauglings,  Arch.  ges.  Physiol.  104,  453-513. 

1908  Tangl,  Franz:    Die    elementare    Zusammensetzung    der    verschiedenen 

Kasei'ne,  Ibid.  121,  534-549. 
1899a  Tarozzi,  G.:  L'acido  fosfocarnico  dei  muscoli  nel    digiuno,    Giornale    d. 

reale  Accad.  di  med.  di  Torino  62,  240-248;  through  Jahresb.  ii.  d. 

Fortschr.  d.  Thierchem.  29  (1899),  460,  461. 
1899b  Tarozzi,  G.:  L'acide    phosphocarnique    des    muscles      dans    le  jeune, 

Archives  ital.  de  biol.  32,  370-378. 
1899    Tauszk,  Franz,  and  Bernhard  Vas:  (Beitrage  zum  Stoffwechsel  bei  Akro- 

megalie),  Pester  med.  -chir.  Presse,  1899,  193-201;  through  Jahrb. 

Neurol.,  1899. 

1904  Taylor,  Alonzo  Engelbert:  Studies  on  an  Ash-free  Diet,  Univ.  of  Cal. 

Pub.  Pathology  1,  71-86. 
1907    Taylor,  Alonzo  Engelbert:  On  the  Synthesis  of  Protein  through  the  Ac- 
tion of  Trypsin,  Jour.  Biol.  Chem.  3,  87-94. 
1911    Taylor,  Alonzo  Engelbert:  On  the  Cutaneous  Elimination  of  Nitrogen, 

Sulfur  and  Phosphorus,  Ibid.  9,  21-24. 
1875    Teissier,  Joseph:  De  la  phosphaturie  a  forme  diabetique,  Lyon  medical 

19,  307-319. 
1877    Teissier,  Louis  Joseph:     Du  diabete     phosphatique.       Recherches     sur 

l'eliminatton  et  variation  des  phosphates  par  les  urines,  These  de 

Paris.      175  pp. 
1883    Tereg  and  Arnold:  Das  Verhalten  der  Calciumphosphate  im  Organismus 

der  Fleischfresser,  Arch.  ges.  Physiol.  32,  122-170. 
1911     Terroine,  E.  F.:  Lecithin  und  diastasische  Wirkungen,  Biochem.  Zeit. 

35,  506;  through  Chem.  Abs.  6  (1912),  2763. 
1910    Teruuchi,  Yutaka:   (Etiology  of  Beri-beri  with  Especial  Reference  to 

Schaumann's  Theory  of  Phosphorus     Starvation),     Saikingakuzashi, 

Tokyo,  1910,  No.  179;  through  Chem.  Abs.  5  (1911),  3111. 
1895    Thezard:  Analyse  d'un  os  de  momie,  Compt.  rend.  Acad  des  sci.  120, 

1126-1128. 

1905  Thompson,  W.  H.,  and  H.  M.  Johnston:  Note  on  the  Effects  of  Pituitary 

Feeding,  Jour,  of  Physiol.  33,  189-191. 
1875    Thudichum,  John  Louis  William:  On  the  Chemical  Constitution  of  the 
Brain,  Chemical  News  31,  112,  113.      . 


696  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1899     Thudichum,  John  Louis  William:  Ueber  den  chemischen  Process  Gallen- 

steinkrankheit  beim  Menschen  und  in  Thieren,  Arch.  path.  Anat.  u. 

Physiol.  156,  384-394. 
1901     Thudichum,  John  Louis  William :  Die  chemische  Konstitution  des  Gehirns 

des  Menschen  und  der  Tiere,  Tubingen.      339+XII  pp. 
1876     Thudichum  and  Kingzett :  Phosphorhaltige  Substanzen  aus  dem  Gehirne, 

Ber.  deut.  chem.  Gesell.  9,  950. 
1885     Tichomiroff,  A.:  Chemische  Studien  iiber  die  Entwickelung  der  Insekten- 

eier,  Zeit.  Physiol.  Chem.  9,  518-532. 

1904  Tigerstedt,  Carl:  Ein  Beitrag  zur  Kenntniss  des  Phosphorstoffwechsels 

beim  erwachsenen  Menschen,  Skand,  Arch.  Physiol.  16,  67-78. 

1911  Tigerstedt,  Robert:  Zur  Kenntniss  der  Aschenbestandteile  in  der  frei- 

gewahlten  Kost  der  Menschen,  Ibid.  24,  97-112. 

1912  Timoshok,  Peter:  Ueber  den  Einfluss  des  Nuclein-natriums  (Merck)  auf 

die  fermentative  Function  der  Organe  und     Gewebe     bei     Staphylo- 
kokkeninfection,  Inaug.  Diss.,  St.  Petersburg,  1912;  through  Zentralbl. 
Biochem.  u.  Biophys.  15  (1913),  762,  763. 
1906    Timpano,  Pietro:  La  cura  fosforata  col  "Protylin,"  La  Clinica  moderna 
12,  565-568. 

1905  Tobler,  Ludwig:  Phosphaturie  und  Calcariurie,  Arch.  exp.  Path.  u.  Ther. 

52,  116-139. 
1911     Tobler,  Ludwig:  Ueber  Veranderungen     im      Mineralstoffbestand      des 
Sauglingskorpers  bei   akuten     und     chronischen     Gewichtsverlusten, 
Jahrb.  Kinderheilk.  73,  566-585. 

1910  Tobler,  L.,  and  F.  Noll:    Zur  Kenntnis    des    Mineralstoffwechsels    beim 

gesunden  Brustkind,  Monatsschr.  Kinderheilk.  9,  210-217. 

1908  Togami,  K.:  Notiz  zur  Kenntnis  des  Phosphorstoffwechsels,  Med.  Klinik, 

Wien.  4,  II,  1837,  1838. 

1897  Tomlinson,  Oscar  R.:  Nuclein  Solution;  a  Clinical  Study,  Jour.  Amer. 

Med.  Assoc.  29,  478,  479. 

1898  Tonelli,   Cesare:   Contributo  clinico   all'   uso   della  fosfoluteina   Serono, 

Gazzett.  degli  ospedali  e  delle  cliniche  19,  I,  710,  711. 

1909  Tornani,  E.:  (Lecithin  and  other  Components  of  Egg  Yolks),  Bollettino 

chim.  farmaceut.  48,  520,  521;  through  Jour.  Chem.  Soc.  96,  II  (1909), 
818. 
1910-11  Towles,  Caroline:  Untersuchungen  iiber  den    Einfluss    des    Phosphor- 
lebertrans  auf  den   Stoffwechsel  eines  rachitischen   Sauglings,  Zeit. 
Kinderheilk.,  Originalien  1,  346-355. 

1911  Toyofuku,  T.:  Ueber  die  parathyreoprive  Veranderung  des  Rattenzahnes, 

Frankfurter  Z.  Pathol.  7,  249-294;  through  Chem.  Abs.  5  (1911),  2397. 

1911  Trier,  Georg:  Aminoathylalkohol,  ein  Produkt  der  Hydrolyse  des  "Leci- 

thins" (Phosphatids)  der  Bohnensamen,  Zeit.  physiol.  Chem.  73,  383- 
388. 

1912  Trier,  Georg:  Ueber  die  Gewinnung  von  Aminoathylalkohol  aus  Eileci- 

thin,  Ibid.  76,  496-498. 
1913a  Trier,  Georg:  Ueber  die  nach  den  Methoden  der  Lecithindarstellung  aus 

Pflanzensamen  erhaltlichen  Verbindungen.      I.  Mitteilung.  Einleitung- 

Bohnensamen,  Ibid.  86,  1-32. 
1913b  Trier,  Georg:  Ueber  die  nach  den  Methoden  der  Lecithindarstellung  aus 

Pflanzensamen  erhaltlichen  Verbindungen.      I.  Mitteilung.  Einleitung- 

gleichende  Hydrolyse  von  Eilecithin,  Ibid.  86,  141-152. 


PHOSPHORUS  METABOLISM  697 

1913c  Trier,  Georg:  Ueber  die  nach  den  Methoden  der  lecithindarstellung  aus 

Pflanzensamen  erhaltlichen  Verbindungen.       III.   Mitteilung.   Hafer- 

samen,  Ibid.  86,  153-173. 
1913d  Trier,  Georg:  Ueber  die  nach  den  Methoden  der  Lecithindarstellung  aus 

Pflanzensamen  erhaltlichen  Verbindungen.       IV.  Mitteilung.   Erbsen, 

Schwarzkiefer,  Reis,  Ibid.  86,  407-414. 
1874     Tripier,  L.:  Recherches  sur  la  production     artificielle     du     rachitisme, 

Archives  de  physiol.  normale  et  pathol.,  (1874)  108. 
1909     Trowbridge,  P.  F.,  and  F.  W.  Woodman:  Changes  in  the  Composition  of 

the  Skeleton  of  Beef  Animals.     (First  paper.)     Jour.  Ind.  and  Engin. 

Chem.  1,  725-732. 
1903     Trunz,  August:  Ueber  die  mineralischen  Bestandteile  der  Kuhmilch  und 

ihre  Schwankungen  im  Verlaufe  einer  Laktationsperiode,  Zeit.  physiol. 

Chem.  40,  263-310. 
1912a  Tschernorutzky,  Helene:  Ueber  das  Vorkommen  von  Nucleinsaure  in 

reifen  Heringseiern,  Ibid.  80,  194-197. 
1912b  Tschernorutzky,  Helene:  Das  Verhalten      einiger      Nucleinsauren      zu 

glukosidspaltenden  Fermenten,  Ibid.  80,  298-306. 

1911  Tschernoruzki,  M.:  Ueber  die  Wirkung  der  Nucleinsaure  auf  die  ferment- 

ativen  Prozesse  im  tierischen  Organismus.  Biochem.  Zeit.  36,  363-375. 

1912  Tschernoruzki,  M.:  Ueber  die  gegenseitige  Wirkung  von  Nucleinsaure 

und  nucleinspaltenden  Ferment  im  tierischen    Organismus,  Ibid.    44, 

353-391. 
1911     Tschikste,  Anastasia:  Ueber  die  Wirkung  des     im     Schilddrusenkolloid 

enthaltenen  Nucleoproteides  bei  Morbus  Basedowii,  Deut.  med.  Woch- 

enschr.  37,  2217-2222. 
1889     von  Tschirwinski,  N. :  Zur  Frage  uber  das  Wachstum  der  Rohrenknochen 

und  den  muthmasslichen  Zusammenhang  dieses  Wachstums  mit  dem 

Wechsel  der  Schneidezahne  bei  den  Schafen,  Landwirtsch.  Jahrb.  18, 

463-475. 
1884     Tuczek,  Franz:     Mittheilungen     von     Stoffweehseluntersuchungen     bei 

abtinirenden  Geisteskranken,  Arch.  Psychiat.  u.  Nervenkrankh.  15,  784- 

799. 
1906    Tunnicliffe,  F.  W.:  The  Behavior  in  the  Body  of  Certain  Organic  and 

Inorganic  Phosphorus   Compounds,   Congres  internat.   de   med.,   Lis- 

bonne,  15,  sect.  4,  181-193  (English);     also    Archives    internat.     de 

pharmacodyn.  et  de  ther.  16,  207-220  (English). 
1906    Tutin,  F.,  and  A.  C.  O.  Hann:  The  Relation  between  Natural  and  Synthet- 
ical Glyeerylphosphorie  Acids.      II.  Jour,  of  the  Chem.  Soc.  89,  1749- 

1758. 
1909    Tyshnjenko,  A.:  (Ueber  den  Einfluss  des  Phytins  und  des  glycerinphos- 

phorsauren  Natriums  auf  den  Stickstoff-  und  Phosphorstoffwechsel.) 

Diss.,  St.  Petersburg.  117  pp.  Through  Jahresb.  u.  d.  Fortschr.  d.  Thier- 

chem.  39  (1909),  598. 
(1911)  Uffenheimer,  A.,  and  Yoshiyiro  Takeno:  Der  Nachweis  des  Kasei'ns  in 

den  sogenannten  Kase'inbrockeln  des  Sauglingsstuhls  mit  Hilfe  der 

biologischen  Methodik,  insbesondere  der  Anaphylaxie,  Zeit.  Kinder- 

heilk.  2,  32-61;  through  Jahresb.  ii.  d.  Fortschr.    d.    Thierchem.    41 

(1911),  298. 
1895    Uhlrich:  Knochenbruchigkeit,  Ber.  ii.  d.  Veterinarwesen  im  Konigreiche 

Sachsen  40,  41. 


698  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1901  Ulpiani,  C:  Attivita  ottica  della  lecitina,,  Atti  d.  reale  Accad.  Lincel, 

ser.  5.,  Rend.  Classe  di  sci.  fis.,  mathemat.  e  natur.  11,  I,  368-375;  421- 
425. 

1902  Ulpiani,  C,  and  G.  Lelli:  Su  un  nuovo  proteide  del  cervello,  Ibid.  11,  II, 

18-24;  also  Gazzetta  chim.  ital.  32, 1,  466-473  (taken  through  Jahresb.  ii 

d.  Fortschr.  d.  Thierchem.  32  (1902),  528,  529). 
1911     Ulrich,  Chr.:  Beitrage  zur  Kenntnis  des  Fischfleisches,  Arch,  der  Phar- 

macie  249,  68-92. 
1896    Umber,  F.:  Ueber  den  Einfluss  nucleinhaltiger  Nahrung  auf  die  Harn- 

saure-bildung,  Zeit.  klin.  Med.  29,  174-189. 

1900  Umber,  F.:  Das  Nucleoproteid  des  Pankreas,  Ibid.  40,  464-479. 

1901  Umber,  F.:  Ueber  die  fermentative  Spaltung     der    Nukleoproteide    ira 

Stoffwechsel,  Ibid.  43,  282-303. 
1895     Umikoff:  Biology  of  Phosphorus,  Diss.,  St.  Petersburg. 

1908  Underhill,  Frank  P.,  and  Tadasu  Saiki:  The     Influence     of     Complete 

Thyroidectomy  and  of  Thyroid  Feeding     upon     Certain     Phases     of 
Intermediary  Metabolism,  Jour.  Biol.  Chem.  5,  225-241. 

1902  Ungar,  E.:  Zur  Phosphorbehandlung  der  Rachitis,  Munch,  med.  Wochen- 

schr.  49,  999-1003. 

1907  Urano,  Fremihiko:  Neue  Versuche  iiber  die  Salze  des  Muskels,  Zeit. 

Biol.  50  (N.  F.  32),  212-246. 
1913     Ushenko,  A.  I.:  (Metabolism  in  Thyroidectomized  Dogs),  Russki  Wratch 

11,  1751-1754;  through  Chem.  Abs.  7  (1913),  3150. 
1910     Usuki:  Die  Fettverdauung  im  Magen  und  Dunndarm  und  ihre  Beein- 

flussung  durch  Lecithin,  Arch.  exp.  Path.  u.  Pharm.  63,  270-293. 

1909  Vageler,   Hans:  Untersuchungen  iiber  das  Vorkommen  von  Phosphat- 

iden  in  vegetabilischen  und  tierischen  Stoffen,  Biochem.  Zeit.  17,  189- 

219. 
1854     Valenciennes,  A.,  and  E.  Fremy :  Recherches  sur  la  composition  des  oeuf s 

dans  la  serie  des  animaux,  Compt.  rend.  Acad,  des  sci.  38,  469-484. 
1905     Valenti,  Adriano:  II  nucleone  nel  latte  di  donna     nei     diversi     periodi 

di  allattamento,  Soc.  med.  -chir.  di  Pavia,  Sitz.  19,  V,  1905;  through 

Biochem.  Centralbl.  4   (1905-6),  550. 

1908  Valenti,  Adriano:  Sul  contenuto  in  nucleone  del  latte  di  donna  durante 

l'allattamento,  Archivio  di  farmacol.  speriment.   7,  447-459;  through 

Jahresb.  u.  d.  Fortschr.  d.  Thierchem.  38  (1908),  284. 
1890-1  Van  Acker  en,  Friedr.:  Ueber  Harnsaureausscheidung  bei  einigen  Krank- 

heiten,  insbesondere  bei  Morbus  Brightii,  Charite-Annalen  17,  206-227. 
1909a  Van  Dam,  W.:  Beitrag  zur  Kenntnis  der  Labgerinnung,  Zeit.  physiol. 

Chem.  58,  295-330. 
1909b  Van  Dam,  W.:  Ueber  die  Wirkung  des  Labs  auf  Paracaseinkalks,  Ibid. 

61,  147-163. 
1912     Van  Dam,  W.:  Die  Verdauung  des  Caseins  durch  Pepsin     vom     Kalb, 

Schwein  und  Rind,  Ibid.  79,  247-273. 
1907    Van  Herwerden,  M.:  Beitrag  zur  Kenntnis  der  Labwirkung  auf  Casein, 

Ibid.  52,  184-206. 
1905     Van  Hoogenhuyze,  C.  J.  C,  and  H.  Verploegh:  Beobachtungen  iiber  die 

Kreatininausscheidung  beim  Menschen,  Ibid.  46,  415-471. 
1900     Vannini,   Giuseppe:  II  ricambio  materiale  nell'   anchilostomoanemia,  II 

Policlinico  7,  Sez.  med.  29-51. 


PHOSPHORUS  METABOLISM  699 

1904    Vannini,  Giuseppe:  Beitrag  zum  Stoffwechsel  bei  Chlorose,  Arch.  path. 

Anat.  u.  Physiol.  176,  375-413. 
(1911)  Vannini,  Giuseppe:  Der  Stoffwechsel  bei  einem  Falle  von  Kalkablager- 

ungen  unter  der  Haut  (Kalkgicht),  Zentralbl.  f.  d.  Physiol,  u.  Path.  d. 

Stoffw.  N.  F.  6,  520-523;  through  Jahresb.  ii.  d.  Fortschr.  d.  Thierchem. 

41,  (1911),  484. 
1914    Van  Slyke,  Donald  D.:  The  Hexone  Bases  of  Casein,  Jour.  Biol.  Chem. 

16,  531-538. 

1911  Van  Slyke,  Donald  D.,  and  George  F.  White:  The  Relation  Between  the 

Digestibility  and  the  Retention  of  Ingested  Proteins,  Ibid.  9,  219-229. 

1907  Van  Slyke,  Lucius  L.,  and  Alfred  W.  Bosworth:  I.  Some  of  the  First 
Chemical  Changes  in  Cheddar  Cheese.  II.  The  Acidity  of  the  Water- 
Extract  of  Cheddar  Cheese,  N.  Y.  Agr.  Exp.  Sta.  (Geneva),  Tech.  Bui. 
No.  4.      22  pp. 

1909  Van  Slyke,  Lucius  L.,  and  Alfred  W.  Bosworth :  A  Volumetric  Method  for 
the  Determination  of  Casein  in  Milk,  Jour.  Ind.  and  Engin.  Chem.  1, 
768-771;  also  N.  Y.  Agr.  Exp.  Sta.  (Geneva),  Tech.  Bui.  No.  10.  20  pp. 

1912  Van  Slyke,  Lucius  L.,  and  Alfred  W.  Bosworth:  Composition  and  Proper- 

ties of  Some  Casein  and  Paracasein  Compounds  and  their  Relations  to 
Cheese,  N.  Y.  Agr.  Exp.  Sta.,  Tech.  Bui.  No.  26.      32  pp. 

1913a  Van  Slyke,  Lucius  L.,  and  Alfred  W.  Bosworth:  Method  of  Preparing 
Ash-free  Casein  and  Paracasein,  Jour.  Biol.  Chem.  14,  203-206. 

•1913b  Van  Slyke,  Lucius  L.,  and  Alfred  W.  Bosworth:  Preparation  and  Com- 
position of  Unsaturated  or  Acid  Casemates  and  Paracaseinates,  Ibid* 
14,  211-225. 

1913c  Van  Slyke,  Lucius  L.,  and  Alfred  W.  Bosworth:  Valency  of  Molecules  and 
Molecular  Weights  of  Casein  and  Paracasein,  Ibid.  14,  227-230. 

1913d  Van  Slyke,  Lucius  L.,  and  Alfred  W.  Bosworth:  Composition  and  Prop- 
erties of  the  Brine-Soluble  Compound  in  Cheese,  Ibid.  14,  231-236. 

1902  Van  Slyke,  L.  L.,  and  E.  B.  Hart:  I.  A  Study  of  Some  of  the  Salts 
Formed  by  Casein  and  Paracasein  with  Acids:  Their  Relations  ta 
American  Cheddar  Cheese,  Amer.  Chem.  Jour.  28,  411-438;  also  N.  Y. 
Agr.  Exp.  Sta.  (Geneva),  Bui.  No.  214,  pp  53-79. 

1904  Van  Slyke,  L.  L.,  and  E.  B.  Hart:  Chemical  Changes  in  the  Souring  of 
Milk  and  their  Relations  to  Cottage  Cheese,  N.  Y.  Agr.  Exp.  Sta. 
(Geneva),  Bui.  No.  245,  36  pp. 

1905a  Van  Slyke,  L.  L.,  and  E.  B.  Hart:  X.  Casein  and  Paracasein  in  Some  of 
their  Relations  to  Bases  and  Acids,  Amer.  Chem.  Jour.  33,  461-496. 

1905b  Van  Slyke,  L.  L.,  and  E.  B.  Hart:  The  Proteids  of  Cream,  Butter  and 
Buttermilk  in  Relation  to  Mottled  Butter,  Jour,  of  the  Amer.  Chem. 
Soc.  27,  679-690. 

1905c  Van  Slyke,  L.  L.,  and  E.  B.  Hart :  Some  of  the  Relations  of  Casein  and 
Paracasein  to  Bases  and  Acids,  and  their  Application  to  Cheddar 
Cheese,  N.  Y.  Agr.  Exp.  Sta.  (Geneva),  Bui.  No.  261.     37  pp. 

1906  Van  Slyke,  L.  L.,  and  D.  D.  Van  Slyke:  I.  On  the  Action  of  Dilute  Acids 

upon  Casein  when  no  Soluble  Compounds  are  Formed.  II.  The 
Hydrolysis  of  the  Sodium  Salts  of  Casein,  N.  Y.  Agr.  Exp.  Sta.,  Tech. 
Bui.  No.  3,  pp  75-160. 

1907  Van  Slyke,  L.  L.,  and  D.  D.  Van  Slyke:  The  Action  of  Dilute  Acids  upon 

Casein  when  no  Soluble  Compounds  are  Formed,  Amer.  Chem.  Jour. 
38,  383-456. 


700  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1894a  Vaudin,  L.:  Sur  l'acide  citrique  et  le  phosphate  de  chaux  en  dissolution 
dans  le  lait,  Annales  de  lTnst.  Pasteur  8,  502-505. 

1894b  Vaudin,  L.:  Sur  le  phosphate  de  chaux  en  dissolution  dans  le  lait,  Ibid. 
8,  856-862. 

1895  Vaudin,  L.:  Sur  le  phosphate  de  chaux  du  lait,  Compt.  rend.  Acad,  des 
sci.  120,  785-787. 

1897a  Vaudin,  L.:  Sur  la  richesse  du  lait  en  elements  mineraux  et  en  phos- 
phates terreux,  Annales  de  lTnst.  Pasteur,  11,  541-544. 

1897b  Vaudin,  L.:  Sur  le  phosphate  de  chaux  du  lait,  La  Laiterie,  51,  52; 
through  Vierteljahreschr.  ii.  d.  Fortschr.  a.  d.  Geb.  d.  Chem.  d.  Nah- 
rungs-  u.  Genussmittel  12  (1897),  167. 

1893  Vaughan,  Victor  C,  Fredk.  G.  Novy,  and  Charles  T.  McClintock:  The 
Germicidal  Properties  of  Nucleins.  A  Preliminary  Contribution  from 
the  Laboratory  of  Hygiene  of  Michigan  University,  Medical  News  62, 
536-538. 

1811  Vauquelin,  M.:  Historique  des  travaux  chimiques  entrepris  jusqu'ici  sur 
la  matiere  cerebrale,  Ann.  of  Philos.  1,  332;  Ger.  trans,  in  Schweigger's 
J.  8   (1813),  430-460. 

1908  Vay,  F.:  Ueber  die  immunisierende  Wirkung  von  Lecithinausziigen  aus 
Pestbazillen,  Deut.  med.  Wochenschr.  34,  II,  2265,  2266. 

1912  Vedder,  E.  B.:  A  Fourth  Contribution  to  the  Etiology  of  Beriberi,  Phil- 
ippine Jour,  of  Sci.,  B,  7,  415-422;  through  Exp.  Sta.  Record  28  (1913), 
764,  765. 

1912  Vedder,  E.  B.,  and  E.  A.  Clark:  A  Study  of  Polyneuritis  Gallinarum.  A 

5th  Contribution  to  the  Etiology  of  Beriberi,  Ibid.  7,  423-462;  through 
Exp.  Sta.  Record  29  (1913),  180. 

1913  Venturi,  F.,  and  V.  Massella :  Influenza  della  Fitina  sulla  eliminazione  dei 

composti  azotuti  in  individuo  normale,  Arch,  di  Farm,  sperim.  e  Sci. 
affini  16,  97-118. 

1849  Verdeil,  F.:  Untersuchung  der  Blutasche  verschiedener  Thiere,  Annalen 
der  Chem.  u.  Pharm.  69,  89-99. 

1900  Ver  Eecke,  A.:  Les  echanges  materiels  dans  leurs  rapports  avec  les 
phases  de  la  vie  sexuelle.  Etude  des  lois  des  echanges  nutritifs  pen- 
dant la  gestation,  Memoires  couronnes  et  autres  memoires  publies  par 
l'Acad.  roy.  de  med.  de  Belgique  15  (1902),  1-159. 

1884  Vermeulen,  Charles  E.  A.:  Onderzoekingen  omtrent  de  physiologische 
werking  van  het  natrium-hypophosphiet,  Doct.  Diss.,  Amsterdam.  49 
pp.  Through  Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  14  (1884),  249- 
252. 

1913  Vernon,  H.  M. :  Die  Rolle  der  Oberflachenspannung  und  der  Lipoide  f  iir  die 
lebenden  Zellen,  Biochem.  Zeit.  51,  1-25. 

1898  Villinger,  A.:  Vom  Phosphor-Stoffwechsel,  Verhandl.  d.  Congr.  f.  innere 
Med.  16,  328-334. 

1902  Vincent,  Swale:  Die  Eiweisskorper  der  glatten  Muskelfasern,  Zeit. 
physiol.  Chem.  34,  417-429. 

1852  Virchow,  Rud.:  Ueber  parenchymatous  Entzundung,  Arch.  path.  Anat. 

u.  Physiol.  4,  261-324. 

1853  Virchow,   Rud:   Das   normale   Knochenwachsthum   und   die   rachitische 

Storung  desselben,  Ibid.  5,  409-507. 
1911     Vivier,  A.:  (Einfluss  der  Fiitterung  mit  Rubenblattern  auf  die  Produc- 
tion und  Zusammensetzung  der  Milch),  Annal.  des  Falsific.  4,  638-641; 
through  Zeit.  Unters.  d.  Nahrungs-  u.  Genussmittel  25  (1913),  61,  62. 


PHOSPHORUS  METABOLISM  701 

1905  Voltz,  W.:  Ueber  den  Einfluss  des  Lezithins  auf  den  Eiweissumsatz  ohne 

gleichzeitige  Asparaginzufuhr  und  bei  Gegenwart  dieses  Amids,  Arch. 

ges.  Physiol.  107,  415-425. 
1880a  Voelcker,  Augustus:  On  the  Composition  of  Ewe's  Milk,  Jour,  of  the  Roy. 

Agr.  Soc.  of  England,  [2]  16,  592-594. 
1880b  Voelcker,  Augustus:  On  the  Composition  of  Goat's  Milk,  Ibid.  [2]  16, 

594,  595. 
1875     Vogt:  Ueber  Wirkung  der  Milchsaure  auf  Knochenwachsthum,  Berlin. 

klin.  Wochenschr.  12,  473. 
1901     Vogt,  Hans:  Ein  Stoffwechselversuch  bei  akuter  Gicht,  Arch.  klin.  Med. 

71,   21-28. 

1906  Vogt,  Hans:  Der  zeitliche  Ablauf  der  Eiweisszersetzung  bei  verschieden- 

er  Nahrung,  Beitrage  z.  chem.  Physiol,  u.  Path.  8,  409-430. 
1877a  Voit,  Carl:  Ueber  das  Verhalten  der  Kalkschalen  der  Huhnereier  bei  der 

Bebrutung,  Zeit.  Biol.  13,  518-526. 
1877b  Voit,  Carl.  Ueber  den  Einfluss  kalkarmen  Futters  auf  die  Knochen,  Amt- 

licher  Ber.  d.  50  Versamml.  deut.  Naturforsch.  u.  Aerzte  in  Miinch- 

en,  242-244. 
1878    Voit,  Carl:  Ueber  den  Einfluss  kalkarmen  Futters  auf  die  Knochen,  Zeit. 

Tiermedizin  4,  128-131. 
1881     Voit,  Carl:  Physiologie  des  allgemeinen  Stoffwechsels  und  der  Ernah- 

rung,  L.  Hermann's  Handbuch  6,  Thl.  1,  79. 
1880    Voit,  Erwin:  Ueber  die  Bedeutung  des  Kalkes  fur  den  thierischen  Organ- 

ismus,  Zeit.  Biol.  16,  55-118. 
1909-10  Voit,  E.,  and  J.  Zisterer:  Bedingt  die  verschiedene  Zusammensetzung 

der  Eiweisskorper  auch  einen  Unterschied  in  ihren  Nahrwert?       II. 

Mitteilung.       Die  physiologische  Wertigkeit  des  Kaseins  und  seiner 

Spaltungsprodukte,  Ibid.  53  (N.  F.  35),  457-498. 
1904    Volhard,  J.:  Wie  wirkt  ein  Ueberschuss  von  kohlensaurem  Kalk  im  Fut- 

ter  auf  die  Ausnutzung  der  Futterbestandteile,  Landwirtsch.  Versuch. 

Stat.  61,  305-312. 
1873    Volkmann,  A.  W.:  Ueber  die  naheren  Bestandteile     der    menschlichen 
.Knochen,  Konigl.  Sachs.  Gesell.  d.  Wissensch.  zu  Leipzig.  Ber.  u.  d. 

Verhandl.  mathemat.  -phys.  Classe  25,  275-305. 
1913    Volter,  B.:  Beitrage  zur  Kenntnis  der  Chemie  der  Krebstumoren,  Biochem. 

Zeit.  55,  260-265. 

1910  Vorbrodt,  Wladyslaw:  Untersuchungen  iiber  die  Phosphorverbindungen 

in  den  Pflanzensamen,  mit  besonderer  Berucksichtigung  des  Phytins, 

Bui.  internat.  de  l'Acad.  des  sci.  de  Cracovie,  Serie  A  1910,  414-511. 
1899    Vosgien  and  Geroline:  Recherches  sur  l'assimilabilite   des  phosphates 

mineraux  et  leur  action  dans  l'alimentation,  Compt.   rend.   Soc.   de 

biol.  [11]  1,  770-772. 
1909    Vozarik,  Am.:  Aciditat,  Ammoniak,  Phosphorsaure  und  Gesamtstickstoff 

im  Kinderharn  bei  eiweissarmer  und  eiweissreicher  Ernahrung,  Arch. 

Kinderheilk.  50,  199-242. 

1911  Vuaflart,  L.:  Azote  et  acide  phosphorique  dans  le  ble  et  dans  la  farine, 

Jour,  d'agric.  pratique  (n.  s.)  22,  395,  396. 

1912  Vuaflart,  L.:  Azote  et  acide  phosphorique  dans  le  ble  et  dans  la  farine, 

Rept.  Eighth  Internat.  Congr.  of  Applied  Chem.  15,  371-374. 
1905a  Wagner,  J.  Ph.:  Le  phosphate  de  chaux  dans  l'alimentation  du  betail  de 
la  ferme,  Jour.  Soc.  cent.  agr.  Belg.  52,  268-281. 


702  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1905b  Wagner,  J.  Ph.:  Le  phosphate  de  chaux  dans  l'alimentation  du  betail  S, 
la  ferme,  Second  Congr.  internat.  de  Paliment.  rationelle  du  betail, 
Rappt,  103-114. 

1906  Wagner,  J.  Ph.:  Le  phosphate  de  chaux  dans  l'alimentation  du  betail  de 
la  ferme,  Bui.  mensuels  Soc.  des  sci.,  agric.  et  arts  de  la  Basse- Alsace, 
40,  100-109. 

1892  Wagner,  Paul  A.  E.:  Ueber  die  diuretische  Wirkung  des  Borax,  Inaug. 
Diss.,  Kiel.      29  pp. 

1902  Waldvogel:  Der  Stoffwechsel  im  Gichtanfall,  Centralbl.  Stoffwechsel-  u. 
Verdauungskrankh.  3,  1-6,  242. 

1906  Waldvogel  and  Tintemann :  Zur  Chemie  des  Jecorins,  Zeit.  physiol.  Chem. 
47,  129-139. 

1877  Walter,  F.:  Untersuchungen  iiber  die  Wirkung  der  Sauren  auf  den  tier- 
ischen  Organismus,  Exper.  Arch.  7,  148. 

1891  Walter,  G.:  Zur  Kenntniss  des  Ichthulins  und  seiner  Spaltungsproducte, 
Zeit.  physiol.  Chem.  15,  477-494. 

1912a  Walters,  E.  H.:  Studies  in  the  Action  of  Trypsin.  I.  On  the  Hydroly- 
sis of  Casein  by  Trypsin,  Jour.  Biol.  Chem.  11,  267-305. 

1912b  Walters,  E.  H.:  Studies  in  the  Action  of  Trypsin.  II.  (a)  On  the  Influ- 
ence of  the  Products  of  Hydrolysis  upon  the  Rate  of  Hydrolysis  of 
Casein  by  Trypsin;  (b)  The  Autohydrolysis  of  the  Caseinates,  Ibid.  12, 
43-54. 

1910  Ward,  Edgar  P.:  Nuclein  by  Intravenous  Injection  in  the  Treatment  of 
Tuberculosis,  Medical  Record  (N.  Y.),  77,  528-531. 

1890  Warschauer,  Eugen:  Ueber  Osteomalacie  und  Untersuchungen  des  Stoff- 
wechsels  bei  derselben,  Inaug.  Diss.,  Wurzburg.      24  pp. 

1908  Waters,  H.  J.:  The  Capacity  of  Animals  to  Grow  under  Adverse  Condi- 

tions, Proc.  Soc.  Prom.  Agr.  Sci.  1908,  XXIX. 

1909  Waters,  H.  J.:  The  Influence  of  Nutrition  upon  the  Animal  Form,  Ibid. 

XXX. 
1909-10  Waters,  H.  J.:  How  an  Animal  Grows,  Kansas  State  Board  of  Agr., 

17th  Biennial  Rept.,  1909-1910. 
1867    Weber,  Otto:  Zur  Kenntniss  des  Osteomalacie,  insbesondere  der  senilen 

und  iiber  das  Vorkommen  von  Milchsaure  in  osteomalacischen  Knoch- 

en,  Arch.  path.  Anat.  u.  Physiol.  38,  1-15. 
1901     Weber,  S.:  Versuche  iiber  kunstliche  Einschrankung  des  Eiweissumsatzes 

bei  einem  fiebernden  Hammel,  Arch,  exper.  Path.  u.  Pharm.  47,  19-47. 
1905    Wechsler,  Emil:  Vorlaufige  Mitteilung    iiber    Ernahrungsversuche    mit 

Phytin  und  Protylin,  Allgem.  Wiener  med.  Zeitung  50,  110,  111. 
1872    Wegner,  Georg:  Der  Einfluss  des  Phosphors  auf  den  Organismus,  Arch. 

path.  Anat.  u.  Physiol.  55,  11-45. 
1876     Wegner,  Rudolph:  Experimentelle  Beitrag  zur  Wirkung  der  Milchsaure- 

injection  auf  die  lebenden  Knochen,  Inaug.  Diss.,  Greifswald. 
1895a  Weintraud,  W.:  Ueber  Harnsaurebildung  beim  Menschen,  Arch.  Anat.  u. 

Physiol.,  physiol.  Abt.,  382-385. 
1895b  Weintraud,  W.:  Ueber  den  Einfluss  des  Nucleins  der  Nahrung  auf  die 

Harnsaurebildung,  Berlin,  klin.  Wochenschr.  32,  405-409. 
1900    Weintraud,  W.:  Ueber  den  Abbau  des  Nucleins  im  Stoffwechsel,  Ver- 

handl.  d.  18.  Congr.  f.  innere  Med.,  232-243. 


PHOSPHORUS  METABOLISM  703 

1911  Weiser,  Stephan:  Ueber  den  Ca-,  Mg-,  P-  und  N-Umsatz  des  ungarischen 

Landschweines,  Mitt,  der  Vers.  -Anstalt  Ungarns  1911,  733-746; 
through  Jahresb.  u.  d.  Fortschr.  d.  Thierchem.  41  (1911),  552. 

1912  Weiser,  Stephan:  Ueber  den  Ca-,  Mg-,  P-  und  N-Umsatz  des  wachsenden 

Schweines,  Biochem.  Zeit.  44,  279-289. 
1871a  Weiske,  H.:  Ueber  den  Einfluss  von  kalk-  und  phosphorsaurearmer  Nah- 

rung  auf  die  Zusammensetzung  der  Knochen.      I.  and  II.,  Zeit.  Biol.  7, 

179-184;  333-337. 
1871b  Weiske,  H.:  Ueber  den  Einfluss  des  als  Beigabe  zum  Futter  gereichten 

phosphorsauren  Kalkes  auf  dem  Aschengehalt  der  Milch,  Annalen  d. 

Landwirtschaft  in  den  Konigl.  Preussisch.  Staaten  11,  309,  310. 
1872a  Weiske,  H. :  Ueber  den  Enfluss  verschiedener  der  Nahrung  beigemengter 

Erdphosphate  auf  die  Zusammensetzung  der  Knochen,  Zeit.  Biol.  8, 

239-245. 
1872b  Weiske,  H.:  Ueber  die  verschieden  Zusammensetzung  des  Ziegenharns 

bei  rein  vegetabilischer  und  rein  animalischer  Nahrung,  Ibid.  8,  246- 

250. 

1873  Weiske,  H.:  Ueber  Assimilation  von  phosphorsaurem  Calcium,  Jour.  f. 

Landwirtsch.  21,  139-154. 

1874  Weiske,  H.:  Ueber  Knochenzusammensetzung     bei     verschiedenartiger 

Ernahrung.     (Vierte  Abhandlung.)     Zeit.  Biol.  10,  410-438. 
1880     Weiske,  H.:  Untersuchungen  iiber  die  Ernahrungs-Vorgange  des  Schafes 

in  seinen  verschiedenen  Altersperioden,  Landwirtsch.  Jahrb.   9,  205- 

300. 
1886     Weiske,  H.:  Ueber  die  Zusammensetzung  von  Blut,  Leber  und  Fleisch 

unter  verschieden  Verhaltnissen,  Jour.  f.  Landwirtsch.  34,  417-  424. 

1888  Weiske,  H.:  Uebt  die  Beigabe  von  neutral  em  phosphorsauren  Calcium  zu 

normal  beschaffenem  Futter  einem  Einfluss  auf  die  Korpergewichts- 
zunahme  der  Thiere  und  auf  das  Gewicht,  resp.  die  Zusammensetzung 
der  Knochen  aus?     Ibid.  36,  279-294 

1889  Weiske,  H.:  Untersuchungen  iiber  Qualitat  und  Quantitat     der     Vogel- 

knochen  und  Federn  in  verschiedenen  Altersstadien,  Landwirtsch.  Ver- 
such.  Stat.  36,  81-103. 

1891a  Weiske,  H.:  Uebt  anhaltende  Aufnahme  von  sauren  Mineralsalzen  einen 
Einfluss  auf  die  Zusammensetzung  der  Knochen  aus?  I.  Ibid.  39, 
17-30. 

1891b  Weiske,  H.:  Uebt  anhaltende  Aufnahme  von  sauren  Mineralsalzen  einen 
Einfluss  auf  das  Korpergewicht  der  Tiere  und  auf  die  Zusammen- 
setzung der  Knochen  aus?       II.  Abhandlung.  Ibid.  39,  241-268. 

1892  Weiske,  H. :  Versuche  iiber  den  Einfluss  welchen  die  Beigabe  verschieden- 

er Salze  zum  Futter  auf  das  Korpergewicht  und  die  Zusammensetzung 
der  Knochen  und  Zahne  ausiibt.      III.  Abhandlung.  Ibid.  40,  81-108. 

1893  Weiske,  H.:  Zur  Beigabe  von  phosphorsauren  Kalk  zum  Futter  der  Tiere, 

Milch-Zeitung  22,  322. 

1894  Weiske,  H.:  Ueber  die  Zusammensetzung     der     Skelette     von     Tieren 

gleicher  Art  und  Rasse,   sowie  gleichen  Alters,  aber  verschiedener 
Grosse,  Landwirtsch.  Versuch.  Stat.  43,  475-480. 
1895a    Weiske,  H.:  Weitere  Beitrage  zur  Frage  iiber  die  Wirkung  eines  Fut- 
ters  mit  sauren  Eigenschaften  auf  den  Organismus,  insbesondere  auf 
das  Skelett,  Zeit.  physiol.  Chem.  20,  595-605. 


704  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1895b  Weiske,  H.:  Zur  Frage  iiber  die  Bedeutung  der  Calciumphosphat-Bei- 
gabe  zum  Futter  f  iir  den  tierischen  Organismus,  Landwirtsch.  Versuch. 
Stat.  45,  242-245. 

1895c  Weiske,  H.:  Versuche  iiber  die  Wirkung  einer  Beigabe  von  Calcium-, 
Strontium-  resp.  Magnesiumcarbonate  zu  einem  kalkarmen,  aber  phos- 
phorsaurereichen  Futter  auf  den  Thierischen  Organismus,  insbeson- 
dere  auf  die  Zusammensetzung  des  Skelettes,  Zeit.  Biol.  31,  421-448. 

1896  Weiske,  H.:  Vergleichende  Untersuchungen  iiber  die  chemische  Zusam- 

mensetzung der  Knochen,  Zahne  etc.  wilder  und  zahmer  Kaninchen, 
Landwirtsch.  Versuch.  Stat.  46,  233-238. 

1897  Weiske,  H. :  Ueber  den  Einfluss  der  Nahrungsentziehung  auf  das  Gewicht 

und  die  Zusammensetzung  der  Organe,  insbesondere  der  Knochen  und 
Zahne,  Zeit.  physiol.  Chem.  22,  485-499. 

1885  Weiske,  H.,  B.  Dehmel,  G.  Kennepohl,  B.  Schulze,  and  G.  Flechsig:  Ver- 
suche iiber  etwaige  Einflusse,  welche  die  Aufnahme  freier  Saure  auf 
die  Verdauungsvorgange  sowie  auf  den  Stickstoff-  und  Mineralstoff- 
Umsatz  im  Korper  der  Herbivoren  ansiibt,  Jour.  f.  Landwirtsch.  33, 
21-76. 

1881  Weiske,  H.,  and  G.  Kennepohl:  Untersuchungen  iiber  Schaf milch  unter 
verschiedenen  Verhaltnissen,  Ibid.  29,  451-472. 

1873  Weiske,  H.,  and  E.  Wildt:  Untersuchungen  iiber  die  Zusammensetzung 
der  Knochen  bei  kalk-  oder  phosphorsaurearmer  Nahrung.  (Dritte 
Abhandlung.)  Zeit.  Biol.  9,  541-549. 

1899  Weiss,  K.:  Ueber  die  Eiweissstoffe  der  Leguminosensamen,  Inaug.  Diss., 
Miinchen.  36  pp.  Through  Jahresb.  u.  d.  Fortschr.  d.  Thierchem.  30 
(1900),  40-42. 

1907  Weiss,  L.:  Beitrage  zur  Kenntniss  der  im  Gerste  und  Malz  vorkommen- 

den  Phosphorverbindungen,  Diss.,  Miinchen.  63  pp.  Through  Jahresb. 
u.  d.  Fortschr.  d.  Thierchem.  37  (1907),  1107,  1108. 

1908  Weissmann,  R.:  Ueber  einige  mit  Phytin  erzielte  Erfolge,  Therapeut. 

Monatsh.  22,  470-472. 

1908  Wellman,  O.:  Untersuchungen  iiber  den  Umsatz  von  Ca,  Mg  und  P  bei 

hungernden  Tieren,  Arch.  ges.  Physiol.  121,  508-533. 
1910    Wells,  H.  Gideon:  The  Purine  Metabolism  of  the  Monkey,  Jour.  Biol. 
Chem.  7,  171-183. 

1909  Wells,  H.  Gideon,  and  Harry  J.  Corper :  The  Purines  and  Purine  Metabo- 

lism of  the  Human  Fetus  and  Placenta,  Ibid.  6,  469-482. 
1905    von  Wendt,  Georg:  Untersuchungen  iiber  den  Eiweiss-  und  Salz-Stoff- 
wechsel  beim  Menschen,  Skand.  Arch.  Physiol.  17,  211-289. 

1909  von  Wendt,  Georg:  Zur  Variability  der  Milch.      2.  Ueber  den  Einfluss 

verschiedener  Salzbeigaben  zum  Futter  auf  Zusammensetzung  und 
Menge  der  Kuhmilch,  Ibid.  21,  89-145. 

1910  von  Wendt,  Georg:  Ueber  den  Einfluss  des  Hohenklimas  auf  den  Stoff- 

wechsel  des  Menschen,  Ibid.  24,  247-258. 
1905    Werner,  Richard:  Zur  Kenntnis  und  Verwertung  der  Rolle  des  Lecithins 

bei  der  biologischen  Wirkung  der  Radium-  und  Rontgenstrahlen,  Deut. 

med.  Wochenschr.  31,  I,  61-63. 
1913    Wesselkin,  N.  W.:  Ueber  die  Ablagerung  vom  fettartigen  Stoffeu  in  dea 

Organen,  Arch.  path.  Anat.  u.  Physiol.  212,  225-235. 
1902    West,  H.  A.:  Therapeutics  of  the  Glycerophosphates,  Merck's  Archivwfc 

4,  263-265. 


PHOSPHORUS  METABOLISM  705 

1882  Weyl,  Th.,  and  H.  Zeitler:  Ueber  die  saure  Reaction  des  thatigen  Muskela 
und  iiber  die  Rolle  der  Phosphorsaure  beim  Muskeltetanus,  Zeit. 
physiol.  Chem.  6,  557-565. 

1903  Wheeler,  Henry  L.,  and  Treat  B.  Johnson:  On  Cytosine  or  2-oxy-6-amino- 
pyrimidine  from  Tritico-Nucleic  Acid,  Amer.  Chem.  Jour.  29,  505-511. 

1913  Wheeler,  Ruth:  Feeding  Experiments  with  Mice,  Jour.  Exp.  Zool.  15, 

209-223. 

1903  Wheeler,  W.  P.:  The  Importance  of  Mineral  Matter  and  the  Value  of 

Grit  for  Chicks,  N.  Y.  Agr.  Exp.  Sta.  (Geneva),  Bui.  No.  242,  293-314. 

1899  White,  W.  Hale,  and  F.  Gowland  Hopkins:  On  the  Excretion  of  Phospho- 

rus and  Nitrogen  in  Leukhaemia,  Jour,  of  Physiol.  24,  42-47. 
1894    Whitfield,  Arthur:  Note  on  the  Chemistry  of  Muscle,  Ibid.  16,  487-490. 
1874    Wibel,  F.:  Die  Constitution  des  Knochenphosphates,  insbesondere  die 

Existenz  und  Bildung  einer  basischen  Verbindung;   (CasPzOs+xCaO). 

Jour.  f.  prakt.  Chem  N.  F.  9,  113-132. 
1912    Wieland,  Hermann:  Beitrage  zur  Aetiologie  der  Beri-Beri.  I.  Analytische 

Untersuchungen  iiber  den  Phosphorgehalt    von     Ernahrungskranken 

Tieren,  Arch.  exp.  Path.  u.  Pharm.  69,  293-306. 

1904  Wilbur,  R.  L.:  Acidosis,  Jour.  Amer.  Med.  Assoc.  43,  II,  1228. 

1900  Wildiers,  Eugene:  Inutility  de  la  lecithine  comme  excitant  de  la  crois- 

sance,  etc.,  La  Cellule  17  (2),  385-405. 

1872  Wildt,  Eugen:  Ueber  die  Zusammensetzung  der  Knochen  der  Kaninchen 
in  den  verschiedenen  Altersstuf  en,  Landwirtsch.  Versuch.  Stat.  15,  404- 
454. 

1874  Wildt,  Eugen:  Ueber  die  Resorption  und  Secretion  der  Nahrungsbestand- 
theile  im  Verdauungskanal  des  Schafes,  Jour.  f.  Landwirtsch.  22,  1-34. 

1879  Wildt,  Eugen:  Studien  iiber  den  Verdauungsprocess  des  Schafes,  Ibid.  27, 
177-248. 

1904  Wiley,  H.  W.,  W.  D.  Bigelow,  and  Others:  Influence  of  Food  Preserva- 
tives and  Artificial  Colors  on  Digestion  and  Health.  I.  Boric  Acid 
and  Borax,  U.  S.  Dept.  Agr.,  Bur.   Chem.  Bui.  No.  84,  Part  I,  477  pp. 

1906  Wiley,  H.  W.,  W.  D.  Bigelow,  F.  C.  Weber  and  Others:  Influence  of  Food 

Preservatives  and  Artificial  Colors  on  Digestion  and  Health.  II.  Sali- 
cylic Acid  and  Salicylates,  Ibid.  84,  Part  II,  479-760. 

1907  Wiley,  H.  W.,  W.  D.  Bigelow,  F.  C.  Weber  and  Others:  Influence  of  Food 

Preservatives  and  Artificial  Colors  on  Digestion  and  Health.  III.  Sul- 
phurous Acid  and  Sulphites,  Ibid.  84,  Part  III,  761-1041. 

1908a  Wiley,  H.  W.,  W.  D.  Bigelow,  F.  C.  Weber  and  Others:  Influence  of  Food 
Preservatives  and  Artificial  Colors  on  Digestion  and  Health.  IV.  Ben- 
zoic Acid  and  Benzoates,  Ibid.  84,  Part  IV,  1043-1294. 

1908b  Wiley,  H.  W.,  W.  D.  Bigelow,  F.  C.  Weber  and  Others:  Influence  of  Food 
Preservatives  and  Artificial  Colors  on  Digestion  and  Health.  V.  Form- 
aldehyde, Ibid.  84,  Part  V,  1295-1500. 

1893  Willdenow,  Clara:  Zur  Kenntnis  der  peptischen  Verdauung  des  Kaseins, 
Inaug.  Diss.,  Bern.      50  pp. 

1904  Willstatter,  Richard,  and  Karl  Ludecke:  Zur  Kenntniss  des  Lecithins, 
Ber.  deut.  chem.  Gesell.  37,  III,  3753-3758. 

1914  Wilson,  D.  H.,  and  P.  B.  Hawk:  Fasting  Studies.      XII.  The  Ammonia, 

Phosphate,  Chloride  and  Acid  Excretion  of  a  Fasting  Man,  Jour.  Amer. 
Chem.  Soc.  36,  137-146. 


706  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1908  Wilson,  R.  A.,  and  W.  Cramer:  On  Protagon:  Its  Chemical  Composition 

and  Physical  Constants,  its  Behaviour  towards  Alcohol,  and  its  Indi- 
viduality, Quarterly  Jour,  of  Exper.  Physiol.  1,  97-110. 
1891    Winkler:  Osteoporose,  Wochenschr.  f.  Tierheilk.  u.  Viehzucht  1891,  No. 
16,  146. 

1905  Winterberg,  Josef:  Ueber  einige  mit  Phj^tin  am  Krankenbette  gesam- 

melte  Erfahrungen,  Aerztliche  Central-Zeitung  17,  406-408. 

1909  Winternitz,  M.  C,  and  W.  Jones:  Ueber  den  Nucleinstoffwechsel  mit  be- 

sonderer  Berucksichtigung  der  Nucleinfermente  in  den  menschlichen 

Organen,  Zeit.  physiol.  Chem.  60,  180-190. 
1897    Winterstein,  E.:  Ueber  einen     phosphorhaltigen     Pflanzenbestandtheil, 

welcher  bei  der  Spaltung  Inosit  liefert,  Ber.  deut.  chem.  Gesell.  30,  II, 

2299-2302. 
1908    Winterstein,  E.:  Ein  Beitrag  zur  Frage  der  Konstitution  des  Phytins, 

Zeit.  physiol.  Chem.  58,  118-121. 

1906  Winterstein,  E.,  and  O.  Hiestand:  Zur  Kenntnis  der  pflanzlichen  Leci- 

thine,  Vorlaufige  Mitteilung.  47,  496-498. 

1908  Winterstein,  E.,  and  O.  Hiestand:  Beitrage  zur  Kenntnis  der  pflanzlichen 

Phosphatide.      II.  Mitteilung.  Ibid.  54,  288-330. 

1909  Winterstein,  E.,  and  K.  Smolenski:  Beitrage  zur  Kenntnis  der  aus  Cer- 

ealien  darstellbaren  Phosphatide.  rV.  Mitteilung.  Ueber  Phosphatide, 
Ibid.  58,  506-521. 

1909a  Winterstein,  E.,  (and  L.  Stegmann):  Beitrage  zur  Kenntnis  pflanzlicher 
Phosphatide.  III.  Mitteilung.  Ueber  ein  Phosphatid  aus  Lupinus 
albus.       (Introduction  by  Winterstein.)  Ibid.  58,  500-505. 

1909b  Winterstein,  E.,  (and  L.  Stegmann) :  Ueber  einen  eigenartigen  phosphor- 
haltigen Bestandteil  der  Blatter  von  Ricinus.  VI.  Mitteilung.  Ueber 
Phosphatide,  Ibid.  58,  527,  528. 

1906  Wintgen,  M.,  and  O.  Keller:  Ueber  die  Zusammensetzung  von  Lecithinen, 

Arch.  f.  Pharmacie  244,  3-11. 
1913     Withers,  W.  A.,  and  B.  J.  Ray:  Studies  in  Cotton  Seed  Meal  Intoxication. 

I.  Pyrophpsphoric  Acid,  Jour.  Biol.  Chem.  14,  53-58;  also  N.  Carolina 

Agr.  Exp.  Sta.  Rept.  1912,  141-145. 
1897    Wittmaack,   Karl:   Ueber  den   Nucleongehalt   der  Kuh-,   Frauen-   und 

Ziegenmilch,  Zeit.  physiol.  Chem.  22,  567-574. 
1900     Worner,  Emil,  and  H.  Thierf elder:  Untersuchungen  iiber  die  chemische 

Zusammensetzung  des  Gehirns,  Ibid.  30,  542-551. 
1903     Wohlgemuth,  Julius:  Ueber  das  Nucleoproteid  der  Leber.  I.  Mittheilung, 

Ibid.  37,  475-483. 
1904a  Wohlgemuth,  Julius:  Ueber  das  Nucleoproteid  der  Leber.  II.  Mittheilung, 

Ibid.  42,  519-523. 
1904b  Wohlgemuth,  Julius:  Ueber  das  Nucleoproteid  der  Leber.  III.  Mitteilung, 

Ber.  deut.  chem.  Gesell.  37,  4362. 
1905     Wohlgemuth,  Julius:  Ueber  das  Nucleoproteid  der  Leber.  IV.  Mitteilung, 

Zeit.  physiol.  Chem.  44,  530-539. 

1907  Wohlgemuth,  Julius:  Untersuchungen  iiber  den  Pankreassaft  des  Men- 

schen.  IV.  Mitteilung.  Ueber  ein  in  ihm  enthaltenes  komplexes 
Hemolysin  und  iiber  die  Darstellung  des  Lecithids,  Biochem.  Zeit.  4, 
271-280;  5,  349. 


PHOSPHORUS  METABOLISM  707 

1911  Wolf,,  Charles  G.  L.,  and  Emil  Oesterberg:  Eiweissstoffwechsel  beim 
Hunde.  II.  Stickstoff-  und  Schwefelstoffwechsel  wahrend  des  Hungers 
und  bei  Unternahrung  mit  Eiweiss,  Kohlenhydraten  und  Fetten,  Ibid. 
35,  329-362. 

1870  Wolff,   Emil:   Die   landw.   -chemische   Versuchsstation   Hohenheim   und 

deren  Thatigkeit  in  den  Jahren  1866-1870.  Berlin.  121  pp.  Autoabstract, 
Landwirtsch.  Jahrb.  2  (1873),  244. 

1871  Wolff,  Emil:  Aschen-Analysen  von     landwirthschaftlichen     Producten, 

Fabrik-Abfallen  und  wildwachsenden  Pflanzen,  Berlin.  194  pp.  About 
2800  analyses. 

1874  Wolff,  Emil:  Die  rationelle  Futterung  der  landwirtschaftlichen'Nutztiere 
(Landwirtschaftliche  Futterungslehre),  Berlin.     239  pp. 

1880  Wolff,  Emil:  Aschen-Analysen  von  land-  und  forstwirthschaftlichen 
Producten,  Fabrik-Abfallen  und  wildwachsenden  Pflanzen,  Berlin.  170 
pp.      About  1600  new  analyses. 

1886  Wolff,  Emil:  Grundlagen  fur  die  rationelle  Futterung  des  Pferdes,  Ber- 
lin.     155  pp. 

1873  Wolff,  E.,  W.  Lunke,  M.  Fleischer,  and  J.  Skalweit:  Versuche  liber  das 
Verdauungsvermogen  von  zweierlei  Schafrassen  in  verschiedenen 
Wachsthumsperioden  und  bei  verschiedener  Futterungsweise,  Land- 
wirtsch. Jahrb.  2,  221-308. 

1912a  Wolff,  G.:  Ueber  den  Kalk-  und  Phosphorsaurestoffwechsel  des  Saug- 
lings  bei  knapper  und  reichlicher  Ernahrung  mit  Kuhmilch,  Inaug. 
Diss.,  Berlin.  29  pp.  Through  Exp.  Sta.  Record  29  (1913),  166. 

1912b  Wolff,  G.:  Ueber  den  Kalk-  und  Phosphorstoffwechsel  des  Sauglings  bei 
knapper  und  reichlicher  Ernahrung  mit  Kuhmilch,  Jahrb.  Kinderheilk. 
76  [3rd  ser.  26],  180-204. 

(1911)  Wolpe,  J.  M.:  Ueber  die  Wirkung  des  organischen  Phosphors  (Phytin) 
beim  runden  Magengeschwur,  Arch.  Verdauungskrankh.  17,  Erganz- 
ungsh.  135-150;  through  Jahresb.  ii.  d.  Fortschr.  d.  Thierchem.  41 
(1911),  280. 

1902  Wood,  Horatio  C,  Jr.:  On  the  Toxic  Action  of  the  Decomposition  Prod- 

ucts of  Lecithin,  Univ.  of  Penn.  Med.  Bui.,  May  1902,  98-101. 

1903  Wood,  Horatio  C,  Jr.:  The  Role  of  Lecithin  in  Pathology  and  Thera- 

peutics, Merck's  Archives  5,  257-259. 
1869     Wood,  Luther  Hodges:  Researches  on  the  Influence  of  Mental  Activity 

upon  the  Excretion  of  Phosphoric  Acid  by  the  Kidneys,  Proc.  of  the 

Conn.  Med.  Soc,  Ser.  2,  3,  197-222. 
1892    Wrampelmeyer,  E. :  Der  Lecithingehalt  der  Butter,  Landwirtsch.  Versuch. 

Stat.  42,  437,  438. 
1894a  Wroblewski,  Augustin:  Beitrage  zur  Kenntniss  des  Frauencasei'ns  und 

seiner  Unterschiede  vom  Kuhcasei'n,  Mittheil.  aus  klinik.  u.  med.  Inst. 

d.  Schweiz  (Annales  suisses  des  sci.  med.)  2,  291-337. 
1894b  Wroblewski,  Augustin:  (Ueber  das  Casein  in  der  Frauenmilch.      Chem- 
ische Unterschiede  zwischen  der  Frauen-  und  Kuhmilch),  Gazeta  lekar- 

ska,  No.  36  and  37,  pp  951  and  988;  through  Jahresb.  ii.  d.  Fortschr.  d. 

Thierchem.  24  (1894),  211-213. 
1898    Wroblewski,  Augustin:  Ein  neuer  eiweissartiger  Bestandteil  der  Milch, 

Zeit.  physiol.  Chem.  26,  308-313. 
1901     Wroblewski,  Augustin:  Ueber  den  Buchner'schen  Hefepresssaft,  Jour,  £, 

prakt.  Chem.  [2]  64,  1-68. 


708  OHIO  EXPERIMENT  STATION:  TECHNICAL  BUL.  5 

1912.  Wuertz,  Ad.:  Versuche  iiber  die  Verteilung  der  Phosphor- Saure  auf 
Harn  und  Kot,  Biochem.  Zeit.  46,  103-111;  through  Chem.  Abs.  7 
(1913),  499. 

1912  Yamigawa,  R.,  T.  Koyana,  H.  Midorikawa  and  T.  Mogi:  Experimentelle 

Studie  iiber  Ursache  und  das  Wesen  von  Kakke,  Mitt.  Med.  Ges.  Tokio 
26,  No.  23,  1-4. 
1884-5  Yeo,  Gerald  F.,  and  E.  F.  Herroun:  A  Note  on  the  Composition  of  Hu- 
man Bile  Obtained  from  a  Fistula,  Jour,  of  Physiol.  5,  116-123. 

1913  Ylppo,  Arvo:  Der  isoelektrische  Punkt  des  Menschen-,  Kuh-,  Ziegen-, 

Hunde-  und  Meerschweinchenmilchcaseins,  Zeit.  Kinderheilk.  8,  Orig- 
inalien,  224-234. 

1910  Yoshimoto,  S.:  Ueber  den  Einfluss  des  Lecithins  auf  den  Stoffwechsel, 

Zeit.  physiol.  Chem.  64,  464-478. 
1907    Young,  William  John:  The  Organic  Phosphorus  Compound  Formed  by 

Yeast- juice  from  Soluble  Phosphates,  Proc.  of  the  Chem.  Soc.  23,  65, 

66. 
1909    Young,  William  John :  The  Hexosephosphate  Formed  by  Yeast- juice  from 

Hexose  and  Phosphate,  Proc.  Of  the  Royal  Soc.  81,  B,  528-545. 

1911  Young,  William  John:  Ueber  die  Zusamensetzung  der  durch  Hefepress- 

saft  gebildeten  Hexosephosphorsaure.      II.  Biochem.  Zeit.  32,  177-188. 

1912  Yushchenko,  A.  I.:  Richesse  de  divers  organes  de  l'homme  et  des  ani- 

maux  en  nuclease,  — ferment  dedoublant  l'acide  nucleique,  Archives  de 

sci.  biol.  17,  1-12;  through  Chem.  Abs.  7  (1913),  1726;  see  also  Jusch- 

tschenko,  1911. 
1879    Yvon,  P.:  Sur  le  lait  phosphate,  Repertoire  de  pharmacie  et  jour,  de  chim. 

med.  7,  403,  404. 
1899    Zadik,  H.:  Stoffwechselversuche  mit  phosphorhaltigen    und    phosphor- 

freien  Eiweisskorpern,  Arch.  ges.  Physiol.  77,  1-21. 
1904    Zaitschek,  Arthur:  Zur  Kenntniss    der    Pepsinsalzsaureloslichkeit    der 

Milch  und  der  Casei'ne,  Ibid.  104,  550-563. 
1901a  Zaky,  Aly:  Influence  de  la  lecithine  sur  l'elimination  de  l'acide  urique, 

Compt.  rend.  Soc.  de  biol.  53,  830-832. 
1901b  Zaky,  Aly:  Sur- l'influence  exercee  par  la  lecithine  sur  l'elimination  de 

l'acide  urique,  Bui.  de  l'Acad.  de  med.  [3]  46,  215. 
1886    Zaleski,  St.  Szcz:  Studien  iiber  die  Leber.      I.  Eisengehalt  der  Leber, 

Zeit.  physiol.  Chem.  10,  453-502. 
1902    Zaleski,  W.:  Beitrage  zur  Verwandlung  des  Eiweissphosphors  in  den 

Pflanzen.       (Vorlaufige  Mitteilung.)       Ber.  deut.  bot.  Gesell.  20,  426- 

433. 
1907    Zaleski,  W.:  Ueber  den  Umsatz  der  Phosphorverbindungen  in  reifenden 

Samen,  Ibid.  25,  58-66. 
1866     Zaleksy:  Ueber  die  Zusammensetzung  der  Knochen  des  Menschen  und 

verschiedener  Thiere,  Hoppe-Seyler's  med.  -chem.  Unters.,  19-48. 
1881     Zander:  Zur  Lehre  von  der  Aetiologie,  Pathogenie  und  Therapie  der 

Rachitis,  Arch.  path.  Anat.  u.  Physiol.  83,  377-391. 
1912    Zaribnicky,  Franz:  Ueber  die  chemische  Zusammensetzung  der  Pferde- 

lymphe,  Zeit.  physiol.  Chem.  78,  327-332. 
1904    Zeiz,  Richard  Herrmann:  Ein  Fall  von  Osteomalacic,  der  trotz  Castra- 
tion und  lange  fortgesetzter  Phosphordarreichung  nicht  zur  Heilung 

kam,  Diss.,  Miinchen.  79  pp. 


PHOSPHORUS  METABOLISM  709 

1895    von  Zeynek,  Rich.:  Chemische  Untersuchung  des  Inhalts  zweier  Lymph- 

cysten,  Zeit.  physiol.  Chem.  20,  462-471. 
1910    Zickgraf :  Ueber  die  Phosphatausscheidung  bei  Tuberculosen  und  Chlor- 

otischen,  Centralbl.  innere  Med.  31,  273-275. 
1909    Ziveri,  Alberto:  Modificazioni  al  metodo  di  ricerca  della  colina  e  nuove 

indagini  sulla  presenza  di  essa  e  di  lecitina  nel  liquido  cefalo-rachideo, 

Rivista  di  patol.  nerv.  e  ment.  14,  134-136. 
1894    Zoja,  Luigi:  Sulla  presenza  di  lecitina  nelle  cellule  alveolari  del  polmone 

e  sul  significato  semeiologico  delle  gocce  mieliniche  dello  sputo,  Gaz- 

zetta  medica,  Torino  45,  801-806;  821-827. 

1912  Zuckmayer,  F.:  Beitrag  zur  Aufnahme  und  Verwertung  von  Kalk  und 

Phosphorsaure  durch  den  Darm,  Arch.  ges.  Physiol.  148,  225-256. 
1905    Zuelzer,  G.:  Ueber  die  klinische  Bedeutung  der  anorganischen  Bestand- 

teile  des  Hams,  Med.  Klinik,  Berlin,  1,  958-960. 
1876    Zuelzer,  W.:  Ueber  das  Verhaltniss  der  Phosphorsaure  zura  Stickstoff 

im  Urin,  Arch.  path.  Anat.  u.  Physiol.  66,  223-251;  282-311. 
1881a  Zuelzer,  W.:  Die  klinische  Bedeutung  der  Phosphorsaure  des  Harns, 

Trans,  of  the  Internat.  Med.  Congr.,  London,  2,  154,  155. 
1881b  Zuelzer,  W. :  Harnuntersuchungen  mit  Rucksicht  auf  off entliche  Hygiene, 

Ibid.  4,  568-570. 
1871    Zum:  Zur  Knochenbriichigkeit  der  Rinder,  Der  Thierarzt  10,  13-16. 
1900    Zuntz,  N.:  Ueber  den  Einfluss  gewisser  phosphorhal tiger  Substanzen  auf 

das  Wachstum,  Therapie  der  Gegenwart  2,  529,  530. 

1913  Zunz,  Leo:  Stoffwechselversuche  bei  Osteomalacic,  Arch.  Gynakol.  99, 

145-166. 
1900    Zweifel,  Paul:  Aetiologie,  Prophylaxis    und    Therapie    der    Rachitis, 
Leipzig.      188  pp.      Through  Congr.  Internat.  de  Med.,  1900,  Sect,  de 
Gynecol.,  583-585. 


INDEX 


Absorption  and  elimination  of  phosphorus,  general,  181 
Absorption  and  secretion  of  phosphorus  in  alimentary  tract,  182,  183 
Absorption  of  calcium  phosphate  as  affected  by  colloidal  state,  185;  colloids, 
185;  intestinal  reaction,  185 
phosphorus  as  affected  by  calcium,  184,  185;  enzymes  of  the 
alimentary  tract,  185,  food  fat-— infants,  459 
Acid  diet,  effects  on  phosphorus  content  of  liver,  136 
Acid  intoxication,  476 
Acidosis,  476 

analytical  measure  of,  477 
from  an  ash-free  diet,  478 
in  diabetes,  510 
in  fever,  479 

influence  on  phosphorus  metabolism,  476 
in  gastrointestinal  disorders  of  children,  478 
symptoms  of,  478 
Acids,  effect  on  bones,  387,  388 

composition  of  flesh,  388 
phosphorus  of  urine,  199 
phosphorus  excretion,  198 
Acromegaly,  in  relation  to  the  hypophysis,  479 

phosphorus  metabolism  in,  479 
Adenase,  definition,  28 
Adenin,  composition,  18 
Adrenals     See  Suprarenal  Capsules 
Age,  influence  of,  metabolism  of  growing  boys,  511,  512 
on  bones,  113 
brain,  133 

the  digestion  of  phytin,  313 
liver,  136 

muscles,  phosphocarnic  acid  content,  305 
nitrogen  and  phosphorus  ratio,  215 
pancreas,  141 
phosphorus  intake,  410 
spleen,  139,  140 
urine,  407 
Alcohol  ingestion,  allantoin  in  urine,  483 

ferment  activity  of  blood  serum,  540 
lecithin  of  liver,  483 
nucleic  acid  metabolism,  240 
phosphatid  content  of  organs,  484 
phosphorus  metabolism,  483 
rate  of  metabolism,  483 
uric  acid  elimination,  483 
Aleuronate,  feeding  experiments,  322,  550 
Alfalfa,  inorganic  phosphorus,  as  affected  by  fertilizer,  99 
phosphorus  in,  as  affected  by  water  supplied,  94 

fertilizer,  99 
hay,  ether  extract  phosphorus,  90 
inorganic  phosphorus,  78 
mineral  analyses,  78,  94 
phosphorus,  78 
Allantoin  in  urine  after  alcohol  ingestion,  483 
"niif*! pit*  spirit    24^    2S1 
of  dogs,  240,  243,  247;  men,  237;  rabbits,  247;  swine,  243 

711 


712  INDEX 

Alloxur  bases,  composition  of,  17 
Alloxuric  bodies.     See  Purins 

Altitude,  influence  of,  nitrogen  and  minerals — men,  429 
Amniotic  fluid,  phosphorus  content,  457 

Anaemia,  effects  of,  on  blood,  496,  499,  507;  cerebrospinal  fluid,  537;  organs, 
507;  phosphorus  and  chlorine  relations  in  the  organs,  508;  urine, 
499;  water  accumulation  in  the  organs,  508 
lecithin  therapy,  501,  529,  531 
nuclein  therapy,  502 
phosphorus  metabolism,  492 
protylin  therapy,  301 
tuberculous,  nuclein-saline  treatment,  586 
Apples,  inorganic  phosphorus  in,  78 
mineral  analyses  of,  78 
phosphorus  in,  78 
Arteriosclerosis,  calcification  from  phosphatid  cleavage,  449 
effects  on  blood,  501 

the  brain,  135 
Arthritis,  excretion  of  phosphorus,  484 
Artificial  food  preparation,  insufficiency  of,  357 
protein-free  milk,  preparation  of,  360 
value  of,  360 
Ash-free  diet,  acidosis  from,  478 

Ash  of  milk  compared  with  the  ash  of  the  young,  167 
Atrophy,  lecithin  therapy,  531 
Autolytic  cleavage  of  phosphatids  and  of  nuclein,  187 

phosphorus  compounds,  187 
Bacteria  in  the  alimentary  tract,  effects  on  nucleoprotein  digestion,  234 
Bananas,  inorganic  phosphorus,  78 
mineral  analyses,  78 
phosphorus,  78 
Barley  bran,  lecithin  phosphorus,  82 
organic  phosphorus,  82 
phosphorus,  82,  84 
grain,  inorganic  phosphorus,  79 
lecithin  content,  87 

(phosphatid)  content,  85,  89 
phosphorus,  79,  82 
organic  phosphorus,  82 
phosphorus,  79,  84,  87 
protein  phosphorus,  79 
kernel,  phosphorus,  84 
Barlow's  disease,  phosphorus  metabolism,  485 
Basedow's  disease,  lecithin  of  blood  serum,  500 
Beans,  lecithin  content,  87 

(brown),  inorganic  phosphorus,  79 

nuclein  and  phosphoprotein  phosphorus,  79 
phosphatid  phosphorus,  79 
phosphorus,  79 
phytin  phosphorus,  79 
(garden),  lecithin  (phosphatid)  content,  85 

phosphorus,  85 
(kidney),  lecithin  (phosphatid)  content,  85 
lecithin  phosphorus,  81 
phosphorus,  81,  85 
phytin  phosphorus,  81 
(navy),  inorganic  phosphorus,  78 
mineral  analyses,  78 
phosphorus,  78 
(Windsor),  inorganic  phosphorus,  79 
lecithin  phosphorus,  79 
phosphorus,  79 
protein  phosphorus,  79 


INDEX  713 

Beechnut  cake,  lecithin  (phosphatid)  content,  85 
Beer,  distribution  of  phosphorus,  80 
Beet  leaves  (sugar),  lecithin  content,  87 

molasses,  favorable  to  phosphorus  absorption,  369 
pulp,  inorganic  phosphorus,  78 
mineral  analyses,  78 
phosphorus,  78 
root  (sugar),  lecithin  content,  87 
Beets  (sugar),  nuclein  and  phosphoprotein  phosphorus,  88 
phosphatid  phosphorus,  88 
phosphorus,  88 
Benzoic  acid,  phosphorus  retention  after,  368 
Beriberi,  cause  of,  484 

lack  of  phosphatese,  491 

phosphorus,  488 
phosphorus  metabolism  disturbance,  485 
Bermuda  grass,  mineral  analyses  as  affected  by  water  supply,  94 
phosphorus  as  affected  by  water  supply,  94 
hay,  ether  extract  phosphorus,  90 
Bile,  general  analyses,  174 

influence  on  lecithin  absorption,  289 
pancreatic  juice,  282 
pancreatic  digestion  of  casein,  270 
inorganic  phosphorus— cattle,  174 

lecithin  content — hedgehog,  107;  men,  174;  rabbit,  107;  several,  174 
mineral  analyses — men,  105 
nucleoproteins — cattle,  174 
phosphorus— men,  105 
phosphorus  content,  173 
Bile  salts  and  lecithin,  solubility  of  fats  and  soaps,  283 
Biocitin,  retention  of  lecithin  from,  295 
feeding  experiments,  295 
lecithin  content,  532 
therapy,  532 
Bioplastin,  description,  93,  531 
Bioson,  composition,  93 

therapy,  532 
Bleeding,  effect  on  composition  of  organs,  507 

Blood  cell  count,  affected  by  acute   pneumonia,  517;    lecithin   ingestion,  533; 
lecithin  injection,  528,  529,  530;  nuclein  therapy  in  blood  diseases, 
502;     osteomalacia,     557;     phytin     ingestion,     533.      See     also 
Leucocytosis 
cells,  differentiation  of  the  phosphatids  of,  153 
circulation  as  affected  by  protylin,  551 

coagulating  action  of  nucleoproteins  and  nucleic  acids,  250,  251,  255 
composition,  affected  by  acromegaly,  480;  anaemia,  496;  arteriosclerosis, 
501;  chlorosis,  496;  crisis  of  diseases,  499;  degen- 
eration and  regeneration  of  severed  nerves,  538; 
diabetes    mellitus,  511;    diet,  369;    diseases,    496, 
498;  lecithin  ingestion,  533;  leukaemia,  495,  497; 
mental    activity,  462;    pain,  463;    pneumonia  and 
anaemia,  507 
lecithin  content,  affected  by  lecithin  injection  or  ingestion, 
498;  nephritis,  548;  polycythaemia  rubra  megalosplenica, 
500 
of  ash,  affected  by  osteomalacia,  557 
partition  of  phosphorus,  affected  by  thyro-parathyroidectomy 

in  the  dog,  560 
phosphatid  content  for  normal  and  pregnant  women  and  for 

new-born  infants,  472 
phosphorus  content,  149 

affected  by    endoarteriitis,    512;    kidney 
diseases,  548 


714  INDEX 

Blood  diseases,  phosphorus  compound  treatment,  501 

metabolism  and  characteristics,  492 
dissolution,  leucocytosis  and  changes  in  metabolism  during,  496 
ether-alcohol  soluble  phosphorus — rabbit,  313 
inorganic  phosphorus  in — cat,  151;  cattle,  152;     dog,  151;     goat,  151; 

horse,  152;  rabbit,  151;  sheep,  152;  swine,  151,  369 
lecithin  content — cat,  151;  cattle,  152;  dog,  151;  goat,  151;  horse,  152; 

men,  498,  500;  rabbit,  107,  151;  sheep,  152;  swine,  151 
mineral  analyses  of — cat,  151;  cattle,  152;  dog,  151;  fowl,  153;  goat,  151; 
horse,  152;    men,  105,  497,  498,  499,  501,  507;     rabbit,  151,  313; 
sheep,  152,  153;  swine,  151,  327,  369 
nuclein  phosphorus — cat,  151;  cattle,  152;  dog,  151;  goat,  151;  horse,  152; 

rabbit,  151;  sheep,  152;  swine,  151 
phosphocarnic  acid — cattle,  150;  dog,  149;  rabbit,  149 
phosphorus— cat,  151;    cattle,   107,  152,  153;    dog,  151,  560;    fowl,  153; 
goat,  151;  horse,  152;  men,  104,  105,  497,  498,  499,  501,  507,  512, 
557;  rabbit,  151,  313;  sheep,  152,  153;  swine,  151,  327,  369 
serum,  effect  of  parathyroidectomy  on  partition  of  phosphorus — dog,  560 
esterase  content  as  affected  by  nervous  diseases,  540 
lecithin  content — men,  542 

as  affected  by  diabetic  lipaemia  and  cholaemia,  501; 
diseases,  500;  epilepsy,  541;  paralysis,  500,  542; 
tabes  and  paralysis,  498;  tabo-paralysis,  498 
normal,  498,  542 
lipase  content  as  affected  by  nervous  diseases,  540 
nuclease  content  as  affected  by  nervous  diseases,  540 
phosphorus  in — dog,  560 

phosphorus  content  in  syphilis  and  cancer,  500 
Bluegrass,  inorganic  phosphorus,  78,  95 

affected  by  fertilizer,  96 
mineral  analyses  of,  78 

affected  by  fertilizer,  96 
phosphorus,  78,  95 

affected  by  fertilizer,  96 
Blue-joint  grass,  mineral  analyses,  affected  by  water  supply,  94 
Body  analyses,  effects  of  phytin  and  sodium  phosphate — rabbit,  313 
Body  composition,  affected  by  disease,  general,  506 
Bone  ash,  constancy  of  composition,  386 

feeding,  compared  with  other  phosphatic  supplements,  828 
influence  on  character  of  bones,  328 

urinary  phosphorus,  206 
phosphorus  balances — swine,  328 
experiments  with  dogs,  427 
diet,  effect  in  experimental  osteomalacia,  394 
formation,  influenced  by  electricity,  390;     parathyroids,  559;     potassium 

salts,  390;  the  thyroids,  578,  579 
katabolism  in  diabetes,  510 

phosphorus  starvation,  394 
on  calcium-  or  phosphorus-poor  diet,  394  . 

marrow,  lecithin  and  phosphorus  content,  118 

lecithin  content — cat,  119;  cattle,  119;     children,  119;     dog,  119; 
hedgehog,  107;  horse,  118,  119;  men,  119,  490,  542;  rabbit, 
m  107,  119;  sheep,  119;  swine,  119 
lecithin  content  as  affected  by  dementia  paralytica,  542 

normal,  542 
phosphorus  content,  affected  by  dementia  paralytica,  542 
phosphorus  in — cat,  119;  cattle,  119;  children,  119;  dog,  119 
horse,  119;  men,  119;  rabbit,  119;  sheep,  119;  swine,  119 
Bone  meal,  inorganic  phosphorus,  78 
lecithin  phosphorus,  82 
phosphorus,  78,  82 
feeding,  nitrogen  and  mineral  balances — dog,  323 


INDEX  715 

Bone  meal,  influence  on  bones,  388,  391 

composition  of  milk,  380,  381 

parts  of  body,  370 
development  of  fowl,  391 
nitrogen  and  mineral  retention,  512 
phosphorus  excretion,  210,  224 
production  of  milk,  381 
mineral  analyses,  78 
retention,  224,  324 
supplement  in  calf  feeding,  421 
Bone  phosphate,  composition,  117 

formula,  118 
Bones,  composition,  affected  by  age,  113 

calcium-poor  diet,  535 
diet,  384 

fast— dog,  440;  rabbit,  441,  443 
lactose  injection,  554 

lecithin,  569;  guinea  pigs,  rabbits,  dogs,  300 
osteomalacia,  554 
rachitis,  575 
tuberculosis,  585 
mineral  matter — swine,  394 

after  treatment  with  glycerin  and  potassium 
hydrate,  113 
relation  of  calcium  and  magnesium,  556 

See  also  Malnutrition  of  the  bones;  Osteomalacia;  Rachitis 
ether-alcohol  soluble  phosphorus — rabbit,  313 
inorganic  phosphorus — dog,  300;  guinea  pig,  300;  rabbit,  300 
lecithin  content — hedgehog,  107;  rabbit,  107 
malnutrition,  534 

mineral  analyses  of — cattle,  112,  113;  children,  572;  dog,  111,  112,  387; 
fowl,  112,  114,  386;  gander,  112;  goat,  112;  goose,  113;  men,  105, 
112,  113;  other  animals,  112;  rabbit,  112,  114,  115,  313,  386,  389, 
390,  441;  sheep,  112,  386,  388;  swine,  327,  394 
phosphorus  in— cattle,  113;  children,  572;  dog,  111,  387;  fowl,  114,  386, 
391;  gander,  112;  goose,  113;  horse,  556;  men,  104,  105,  113,  462, 
556;  rabbit,  114,  115,  313,  386,  389,  390,  554-555;  rats,  506;  sheep, 
386,  388;  swine,  327,  394 
Boric  acid,  phosphorus  retention,  368 

Brain,  composition,     affected    by  diseases;     anaemia,     507;     carcinoma,  507; 

dementia  praecox  and  general  paralysis, 
540;  epilepsy,  541;  paralysis,  539;  pneu- 
monia, 507 
growth,  133 

lecithin— guinea  pigs,  rabbits,  dogs,  300 
phosphatids  affected  by  age,  135,  136 

alcoholism,  484 
phosphocarnic  acid  content,  305,  446 
phosphorus  .compounds,  affected  by  tuberculosis,  588 
normal,  588 

under  various  conditions,  132 
content,  affected  by  age,  135;     arteriosclerosis,  135;     nerve 
degeneration,  542;  polyneuritis,  490;  tubercu- 
losis, 588 
normal,  588 
Thudichum's  classification  of  the  constituents,  128 
Brain  and  nerves,  cephalin  phosphorus — guinea  pigs,  588 

ether-alcohol  soluble  phosphorus — rabbit,  313 
extractive  phosphorus — children,  132;  men,  132;  dog,  132 
inorganic  phosphorus — dog,  134 

lecithin  content— cat,  107;  children,  110,  111;  dog,  130,  300; 
guinea  pigs,  300;  hedgehog,  107;  men,  130,  539;  new-born 
infants  and  fetuses,  110,  111,  130;  rabbit,  107 


716  INDEX 

Brain  and  nerves,  lecithin  phosphorus — guinea  pigs,  588 
lecithins  and  cephalins — children,  133 

lipoid  phosphorus — men,  132;  dog,  132,  134;  children,  132,133 
mineral  analyses — men,  105,  507;  rabbit,  313;  swine,  327 
nuclein  content — men,  130 

phosphorus — dog,  132 
nucleoprotein  content — men,  130 
nucleoproteins — dog,  130 
phosphatid  content— dog,  135;  new-born  infants  and  fetuses, 

135 
phosphatids — guinea  pigs,  588 

phosphocarnic  acid — cat,  130;  cattle,  130;  dog,  130,  305;  fowl, 

130;  guinea  pig,  130;  men,  130;  rabbit,  130;  sheep,  130; 

swine,  130 

phosphorus  in — cattle,  107,  131;  children,  132,  133,  134;  dog, 

132,  134,  300;  new-born  infants  and  fetuses,  134,  135; 

guinea  pigs,  300,  588;  horse,  131;  men,  104,  105,  129,  131, 

462,  507,  539;  rabbit,  300,  313;  sheep,  149;  swine,  131,327 

protein  phosphorus — children,  132,  133;  dog,  132;  men,  132 

Bran,  compared    with    acid-washed    bran,  metabolism    and  milk  production 

experiments — cows,  306 
phosphatic  supplements,  balance  and  slaughter  experi- 
ments— swine,  328 
washed  bran  and  calcium  phosphates,  maintenance  and 

slaughter  experiments — swine,  327 
washed  bran  and  washed  bran  plus  potassium  phytate, 
balance  and  milk  production  experiments— cow,  312 
influence  on  character  of  bones,  328 

composition  of  parts  of  the  body,  327,  370 
phosphorus  metabolism,  326,  487 
nuclein  and  phosphoprotein  phosphorus,  88 
phosphatid  phosphorus,  88 
phosphorus,  88 
Bread,  inorganic  phosphorus,  77 
lecithin  content,  87 
mineral  analyses,  77 

nuclein  and  phosphoprotein  phosphorus,  88 
phosphatid  phosphorus,  88 
phosphorus,  77,  88,  366 

white  and  whole  wheat,  nitrogen  and  phosphorus  balances — men,  366 
nitrogen  and  phosphorus  content,  366 
nutritive  value — man,  367;  pigeon,  489 
Brewer's  grains,  lecithin  content,  87 

products,  phosphorus  compounds  of,  80 
Bromlecithin  in  anaemia,  502 

chlorosis  and  secondary  anaemia,  501 
Bronchopneumonia,  effects  on  urinary  phosphorus,  516 
Buckwheat  grains,  lecithin  (phosphatid)  content,  85 
Buffalo  grass,  ether  extract  phosphorus,  90 
Bunge's  Law,  167 

Burr  clover,  ether  extract  phosphorus,  90 
Butter,  contains  a  component  essential  to  normal  growth,  358,  362 

phosphorus,  162 
Buttermilk,  lecithin,  164 
Cabbage,  inorganic  phosphorus,  78 
mineral  analyses,  78 

nuclein  and  phosphoprotein  phosphorus,  88 
phosphatid  phosphorus,  88 
phosphorus,  78,  88 
Cachexia,  lecithin  therapy,  528 
Calcareous  degeneration,  cause,  503 


INDEX  717 

Calcium  carbonate,  fed  with  hay,  effect  on  digestion,  223 

oats,  389 

skim  milk,  mineral  balances — calves,  222 
influence  on  amount  and  composition  of  goat  milk,  383 
bone  katabolism,  384 
calcium  and  phosphorus  retention,  376 
growth,  228 

phosphorus  absorption,  548 
phosphorus  elimination,  203,  209,  222,  385 
phosphorus  storage,  425 
Calcium  in  relation  to  rachitis,  567 

loss,  affected  by  hypophysis  injection  in  fast,  482 

in  acromegaly,  480;  diabetes  mellitus,  511 
of  bone,  replaced  by  magnesium  and  strontium,  386 
phosphate,  effects  on  growth  of  offspring,  227 

precipitated,  phosphorus  balances — swine,  328 
use  of,  with  beet  leaves,  225 
See  also  Bone  Meal  and  Bone  Ash 
Calcium-poor  diet,  effects  on  bones,  554;  tissues,  225 

limits  phosphorus  storage  from  corn,  425 
nitrogen  and  mineral  balances — horse,  423 
Calcium  salts,  effect  on  bone  formation,  225 

bones,  385,  386,  389,  391;  tissues,  226 
relation  to  phosphorus  absorption,  548 

excretion,  198,  203,  209,  211,  221,  385 
metabolism.         See     also     Phosphaturia, 

Rachitis  and  Calcium  Metabolism 
retention,  217,  218,  220,  224,  225,  376,  425 
Cancer,  effect  on  composition  of  the  blood,  500;  liver,  504;  organs,  507 
phosphorus  studies  in  connection  with,  503 
presence  of  nuclease,  504 

sodium  nucleate  injection  before  operations  for,  504 
Carbohydrate-phosphoric  esters,  feeding  experiments,  550 
Carbonophosphates,  definition,  154 

estimations  of — milk,  154 
in  milk,  treatments  affecting,  154 
Carbonates,    alkaline    earth,    supplements    to     calcium-poor,  phosphorus-rich 

rations,  390 
oats,  389 
Carbohydrates,  constituents  of  nucleic  acids,  19 
Caries,  504 

fungosa,  lactic  acid  treatment,  504 
Carnaubon,  chemistry,  71 

occurrence,  142 
Cartilage,  phosphorus  compounds,  120 
Casein-calcium  preparations,  treatment  of  rachitis,  576 
Casein,  chemistry,  32 

cleavage  by  colon  bacillus,  269 
feces,  269 
products  of,  hydrolytic,  41 
coagulation,  by  acids  and  by  heat,  36 
renin,  38 

Bang's  description,  40 
Bosworth's  interpretation,  40 
conditions  favorable  to,  38 
the  enzyme,  38 

Hammarsten's  interpretation,  39 
in  the  stomach,  261 
Mellanby's  interpretation,  41 
Schryver's  interpretation,  41 
composition,  elementary  analysis  and  empirical  formula,  34 
relation  of  phosphorus  to  the  molecule,  43,  44 


718  INDEX 

Casein  content  of  cream,  163,  164 

fermented  milks,  162 

milk,  affected  by  fat  in  the  diet,  379 

general  state  of  nutrition,  379 
pathological  states,  382 
relation  to  other  constituents,  153 
skimmed  milk,  163,  164 

whole  milk,  155,  156,  157,  161,  163,  164,  166,  167 
digestion,  261 

feeding  experiments,  272,  293,  427 

See  also  Casein  maintenance  experiments 
in  vitro,  185,  267 
peptic,  affected  by  carbohydrates,  267 

degree  of  acidity,  268 
Salkowski's  description,  264 
products  of  peptic,  263,  264 
rectal  feeding  experiment,  404 
differences  in,  from  different  species,  43,  265 
different  species,  peptic  digestion,  267 
effect  on  ferratin  of  liver,  301 

flow  and  composition  of  milk,  383 
in  feces  of  infants,  458 

maintenance  experiments,  320,  326,  331,  342,  343,  359,  360,  362,  363,  454 
occurrence  of,  33 

physical  and  chemical  properties,  35 
products  of  partial  cleavage,  value  as  food,  272,  356 
racemization  of,  36 
secretion,  260 
Caseonphosphate  of  calcium,  treatment  of  rachitis,  576 
Castration  and  ovariotomy,  phosphorus  metabolism,  559 

metabolism  and  composition  of  the  body,  506,  580 
Cattle,  mineral  analyses  of,  106 

phosphorus  in,  106 
Caudol,  description,  93 
Cephalin,  chemistry,  67 

composition,  elementary  analyses,  empirical  formulae,  68 
fatty  acid  component,  67 
nitrogenous  base,  67 
estimations  of — organs,  588 
feeding,  effect  on  phosphorus  of  brain,  303 
in  intermediary  metabolism,  278 
occurrence,  67 

of  brain  and  nerves,  special  studies,  130 
organs  as  influenced  by  tuberculosis,  588 
Cereal  foods,  mineral  metabolism — swine,  375 

affected  by  calcium —  swine,  375 
Cerebrospinal  fluid,  changes  in  nerve  degeneration,  536 

nucleoprotein  content,  general  paralysis,  542 
■Chestnuts,  lecithin  (phosphatid)  content,  85 

phosphorus  in,  85 
Chickens,  ether-soluble  phosphorus,  448 
inorganic  phosphorus,  448 
phosphorus,  448 
vitellin  phosphorus,  448 
Children,  mineral  analyses,  111 

phosphorus,  111 
Chlorosis  and  tuberculosis  compared  as  to  phosphorus  and  chlorine  elimination, 
584 
bromlecithin  treatment,  502 
influence  on  composition  of  blood,  496 

blood  serum  and  erythrocytes,  498 
phosphorus  metabolism  and  other  characteristics,  492 


INDEX  J19 


Chlorosis,  lecithin  treatment,  530,  532 

nuclein  treatment,  effect  on  blood  counts,  502 
Choline,  constitution,  60 

in  nerve  degeneration,  536 
Chorea,  lecithin  therapy,  532 
Chyle,  lecithin  content,  177 

phosphorus,  177 
Chymosin,  identity  with  pepsin,  38 

occurrence  and  action,  38 
Clover,  phosphorus  affected  by  amount  of  water  supplied,  93 
hay,  inorganic  phosphorus,  78 
mineral  analyses,  78 
phosphorus,  78 
Cocoanut  cake,  lecithin  (phosphatid)  content,  85 
Colamin,  mother  substance  of  choline,  61 
Corn  bran,  phosphorus,  83 
Corn,  effect  on  composition  of  bones,  391 
germ,  phosphorus,  83 
gluten,  phosphorus,  83 
grains,  inorganic  phosphorus,  77,  79 

lecithin  (phosphatid)  content,  85,  87 

phosphorus,  79 
mineral  analyses,  77 
phosphorus,  77,  79,  83,  87 
protein  phosphorus,  79 
nature  of  the  phytin,  53 
shucks,  ether  extract  phosphorus,  90 
specific  effects  on  swine,  370,  376,  425 
stover,  inorganic  phosphorus,  78 
mineral  analyses,  78 
phosphorus,  78 
Cottonseed  cake,  lecithin  phosphorus,  91 

nuclein  phosphorus,  90,  91 
phosphorus,  91 
Cottonseed  meal,  inorganic  phosphorus,  78 
mineral  analyses,  78 
nature  of  the  phytin,  53 
phosphorus,  78 
toxic  effects  of,  91,  226,  227 
Cowpea  hay,  ether  extract  phosphorus,  90 
inorganic  phosphorus,  78 
mineral  analyses,  78 
phosphorus,  78 
Cowpeas,  inorganic  phosphorus,  78 
mineral  analyses,  78 
phosphorus,  78 
Cream,  casein  content,  163,  164 

glycerophosphate  phosphorus,  163 
lecithin  phosphorus,  163 
organic  phosphorus,  163 
m  phosphorus,  163,  164 
Cretinism  attributed  to  hypophysis  and  thyroid  glands,  481 
metabolism  studies,  578 
phosphate  treatment,  579 
thyroid  treatment,  546,  579 
Cuorin,  chemistry,  68 

occurrence,  125,  126,  142 
Cytosin,  composition,  19 
Dates,  inorganic  phosphorus,  78 
mineral  analyses,  78 
phosphorus,  78 
Deaminases,  definition,  28 


720  INDEX 

Degeneration  and  regeneration    of  severed   nerves,  chemical   changes  of  the 

blood  accompanying,  538 
Degenerative  nervous  diseases,  organic  phosphorus  in  the  urine,  539 
Dementia  paralytica,  effect  on  cerebrospinal  fluid,  537 
praecox,  effects  on  brain,  540 

effect  on  ferment  activity  of  cerebrospinal  fluid,  541 
Demineralization,  phytin  treatment,  533 

Dental  tartar  as  derived  from  carbonophosphates  of  the  saliva,  176 
Dentine  of  normal  and  diseased  teeth,  composition,  504 
Development  of  young  as  influenced  by  diet  of  mother,  373 
Diabetes,  glycerophosphate  treatment,  521 

mellitus,  composition  of  blood,  erythrocytes  and  blood  plasma,  498 
lecithin  content  of  blood  serum,  500 
phosphorus  metabolism,  510 
phosphaturia  in.     See  Phosphaturia,  561 
Diabetic  acidosis,  calcium,  magnesium  and  phosphorus  loss,  478 

lipaemia,  effect  on  lipoids  of  blood  serum,  501 
Diamino-monophosphatids,  chemistry,  69 
Diastase,  influence  of  lecithin,  285 

lipoids,  286 
Diet  and  species,  effects  on  phosphorus  elimination,  207 
Dietaries,  inorganic  constituents,  409,  410 

Digestion  and  absorption  of  lecithin  and  glycerophosphates,  286 
influence  of  phosphates,  186 
of  fat,  related  to  lecithin  content  of  bile,  173 

nucleic  acids  and  nucleoproteins,  animal  experiments,  237 
nUcleoproteins  by  enzymes  from  alimentary  tract,  231 
Digestive  mucosa,  phosphorus  compounds,  relation  to  food  digestion,  146 

secretions,  phosphorus  content,  172    _    . 
Diphtheria,  phosphorus  and  nitrogen  in  the  urine,  516 
toxin,  effect  on  composition  of  heart,  583 
metabolism,  583 
Diseases,  general  consideration,  composition  of  the  body,  507 
Diuresis  from  inorganic  salts,  546 
Dogs,  phosphorus,  581 

Dwarf  state  due  to  hypofunctioning  of  hypophysis,  482 
Dyspepsia,  effects  on  phosphorus  retention,  459 
Dyspnoea,  experimental,  effect  on  phosphorus  elimination,  479 
Earth  phosphates  of  the  urine  in  pneumonia  and  typhoid  fever,  519 
Eck  fistula,  definition,  239 

Edestin  feeding  experiments,  321,  322,  326,  331,  333,  431 
Eggs,  changes  during  incubation,  276,  446 
feeding  experiments,  323 
fish,  nucleic  acid  content,  171 
nucleoalbumins,  170 
phosphorus  compounds,  148 
fowl,  ether-soluble  phosphorus,  448 

inorganic  phosphorus,  78,  441,  448 
lecithin  content,  87,  447 
mineral  analyses,  78,  169-170,  441 
nucleoalbumins,  170 
phosphorus,  78,  169,  441,  448 
variability  in  composition,  170 
vitellin  phosphorus,  448 
frog,  organic  phosphorus  compounds,  147 
nutritive  value  as  sole  diet  of  men,  291 
phosphorus,  169 

sea-urchin,  phosphorus  changes  during  development,  449 
Egg  white,  mineral  analyses,  169 

nuclein  and  phosphoprotein  phosphorus,  88 
phosphatid  phosphorus  content,  88 
phosphorus,  88 


INDEX  721 

Egg  yolk,  differentiation  of  phosphatids,  171 
effect  on  digestion  of  milk,  533 

lipoid  deposits,  302 
lecithin  content,  171 
mineral  analyses,  170 

nutritive  value,  compared  with  that  of  milk,  290,  291 

plasmon,  291 
treatment  in  anaemia  and  infantile  athrepsia,  501 
polyneuritis,  489 
Elements  involved  with  phosphorus  in  katabolic  processes,  213;  calcium  and 

magnesium,  203,  377,  575;  nitrogen,  215;  sodium  and  potassium,  201 
Elimination  of  phosphorus,  cutaneous,  184 

•  factors  influencing  the  paths,  182;  acids,  198;  acids 
of  fermentation,  459;    bone  dust,  512;     calcium 
and  magnesium,  203,  377,  574;  diet  of  infants, 
418;  fats,  478;  indigestion  of  breast-fed  infants, 
457;  rickets,  573-574;  salts  of  sodium  and  potas- 
sium/ 201;  species,  221;  summary,  211;  type  of 
diet  and  species,  207 
relation  to  nitrogen  elimination,  214 
phosphates  in  the  urine,  method,  188 
Endogenous  and  exogenous  purins,  definition,  246 

methods  of  estimation,  246 
purin  elimination,  factors  influencing,  246 
Endoarteriitis,  nitrogen,  calcium,  magnesium  and  phosphorus  balances — men, 
512 
phosphorus  content  of  blood — men,  512 
metabolism,  512 
Enzyme  activities,  affected  by  administration  of  lecithin,  281 

nucleic  acids,  252 
phosphates,  186 
of  blood  serum,  effect  of  nervous  diseases,  540 
cerebrospinal  fluid  in  nervous  diseases,  540 
organs  as  affected  by  nucleic  acid,  250 

phosphorus  of  diet,  343 
Enzymes,  casein-coagulating,  occurrence,  38 

effecting  cleavage  of  phosphorus  compounds,  185 
of  individual  organs.    Nuclein  ferments,  235 
reversible  action,  266 
Epilepsy,  effect  on  composition  of  brain,  541 

ferment  activity  of  blood  serum,  540 

cerebrospinal  fluid,  541 
lecithin  of  blood  serum,  541 
phosphorus  content  of  cerebrospinal  fluid,  537 
metabolism,  541 
choline  as  a  cause,  537 

organic  phosphorus  in  urine  after  attack,  541 
phosphorus  treatment,  547 
Erepsin,  digestion  of  nucleic  acids,  232 
Esterase  of  blood  serum  in  nervous  diseases,  540 
Etherization,  delayed  phosphorus  elimination  after,  526 
Eucasein,  description,  92 
Exercise,  influence  on  decomposition  of  bone  tissue,  406 

nitrogen,  calcium  and  phosphorus  balances,  413 
phosphorus  metabolism,  464 
ratio  of  nitrogen  to  phosphorus,  469 
uric  acid  elimination,  466,  469 
urinary  phosphorus,  461 
nuclein  katabolism  of,  reduced  by  training,  466 
Exophthalmic  goiter.    See  Morbus  Basedowii 


722  INDEX 

Fast,  influence  on  composition  of  parts  of  body,  305,  438 
phosphorus  metabolism,  438,  481,  512 

Summary  of  evidence,  446 
phosphorus  excretion,  214,  399,  426 
protein  katabolism,  217 
Fats  and  soaps,  solubility,  affected  by  bile  salts  and  lecithin,  283 
Fatty  degeneration  of  liver,  glycerophosphoric  acid  of  urine,  192 

effect  on  lecithin  of  liver,  513 
Feathers,  mineral  analyses,  127 

phosphorus  content,  126 
Feces,  autoputref action  of  nuclein,  234 
lecithin,  288 
mineral  analyses,  105 
phosphorus  content,  105,  208 

and  compounds,  194 
See  also  Phosphorus  balances  and  phosphorus  excretion 
purins,  due  to  bacteria,  234 
Fermentation  of  sugars,  influence  of  phosphates,  31 
Ferments.     See  Enzymes 
Ferratin,  occurrence,  138 
Fersan,  composition,  93,  550 

feeding  experiments  with  geese  and  men,  550 
Fertilizers,  effects  on  composition  of  foods,  95 
Fetuses  and  new-born  young,  phosphorus  content,  104,  108,  471 
Fever,  acidosis,  479 

experimental,  gas  exchange,  516 
phosphorus  metabolism,  514 
scarlet,  sodium  nucleate  treatment,  519 
Fish,  lecithin  (phosphatid)  content,  122 
mineral  analyses,  121 
phosphorus,  121,  122 

phosphorus,  calcium  and  magnesium  retention  after,  368 
Fistula,  Eck,  definition,  239 

use  in  metabolism  experiments,  239 
London's,  in  nucleic  acid  studies,  244 
Flax  seeds,  lecithin  (phosphatid)  content,  85 

phosphorus,  85 
Floats,  balance  and  slaughter  experiments  with  swine,  328 

See  also  Rock  Phosphate 
Flour,  graham,  inorganic  phosphorus,  79 

nuclein  and  phosphoprotein  phosphorus,  79 
phosphatid  phosphorus,  79 
phosphorus,  79 
phytin  phosphorus,  79 
red  dog,  mineral  analyses,  77 
rice,  inorganic  phosphorus,  79 

nuclein  and  phosphoprotein  phosphorus,  79 
phosphatid  phosphorus,  79 
phosphorus,  79 
phytin  phosphorus,  79 
rye,  inorganic  phosphorus,  79 

nuclein  and  phosphoprotein  phosphorus,  79 
phosphatid  phosphorus,  79 
phosphorus,  79 
phytin  phosphorus,  79 
wheat,  factors  influencing  phosphorus  content,  80,  99 
germ,  nuclein  phosphorus,  78 
phosphorus,  78 
phytin  phosphorus,  78 
inorganic  phosphorus,  77 
mineral  analyses,  77 
phosphorus,  77,  78,  80,  99 
value  affected  by  milling  process,  585 


INDEX  723 

Food  phosphorus,  advantage  of  complexity  of  organization,  355 
Foods,  composition,  affected  by  fertilizers,  95 

in  relation  to  phosphorus  metabolism,  Sbb 
lecithins,  84 
nuclein  phosphorus,  90 
phosphorus  compounds,  76 
phytin,  81 

references,  total  phosphorus  content,  103 
Formaldehyde,  phosphorus  retention  after,  368 
Gastric  juice,  phosphorus,  172;  dog,  172;  men,  199 

phosphate  content  dependent  on  degree  of  acidity,  200 
Genitalia,  organic  phosphorus— ass,  148;  cattle,  148;  fish,  147;  hedgehog,  107; 
horse,  148;  rabbit,  107 
phosphorus— cattle,  149;  fish,  146,  147 
phosphorus  changes  with  season,  147 
content,  146 
Giantism  due  to  excessive  functioning  of  hypophysis,  482 
Glidine,  components,  533 

feeding  experiments,  533 
Glucosides,  digestion,  233  \  ■ 

Glycerophosphate  and  lecithin,  physiological  effects,  bZZ,  58Z 
of  calcium,  commercial,  constituents,  74 

casein  (sanatogen),  feeding  experiments,  325 
Glycerophosphates,  acid  and  neutral  salts,  physiological  effects,  520 
effect  on  ferratin  of  liver,  301 

intracellular  ferments,  281 
phosphorus  excretion,  209,  521 
power  of  oxidation,  280 
respiration  coefficient,  280 
metabolism  of,  digestion  and  absorption,  286 

feeding  and  injection  experiments,  209,  324, 

325,  342,  343-354,  520,  582 
See  also  Glycerophosphoric  acid  compounds 
specific  effects  on  swine,  343-354 
synthesis,  74 
urine,  190 
Glycerophosphate  therapy,  518,  520,  546,  582 

with  antirabes  treatment,  521,  583 
Glycerophosphoric  acid  compounds,  balance  experiments  with,  290 

growth  and     composition     of     animals  as 

affected  by,  296 
metabolism,  276 
See  also  Glycerophosphates 
cream,  163 

decomposition  in  the  organs,  520 
elimination,  path,  209 
in  the  blood,  520 

urine  in  fatty  degeneration,  513 
of  urine,  190 
skimmed  milk,  163 
whole  milk,  163,  166 
Gout,  as  disturbance  of  purin  metabolism,  525 
associated  with  phosphaturia,  565 
phosphorus  metabolism,  522 

purin  excretion  on  light  and  dark  meat  compared,  525 
Grass  (Guam),  ether  extract  phosphorus,  90 

mineral  analyses,  as  affected  by  fertilizer,  95 
(Para),  ether  extract  phosphorus,  90 
phosphatid  phosphorus,  89 
phosphorus  as  affected  by  fertilizer,  95 

See  also  Bermuda,  buffalo,    blue-joint,  Indian  bunch,    Johnson  and  blue 
grass;  also  meadow  fescue 


724  INDEX 

Growth  and  composition  of  animals  as  affected  by  compounds  of  glycerophoa- 
phoric  acid,  296 
affected  by  inorganic  phosphates,  227 
effect  on  phosphorus  requirement,  389 
Guanase,  definition,  28 
Guanin,  composition,  18 
Guanylic  acid,  composition,  22 

digestion,  233,  236 
effects,  250 
Haematogen,  Bunge's  theory,  46 
nature,  172 

phosphorus  content,  46 
Hard  water,  influence  on  teeth,  394 
Hay  and  grass  compared  in  feeding  experiments,  373 
compared  with  oats  in  effect  on  bones,  389 
mineral  analyses,  422 
phosphatid  phosphorus,  89 

phosphorus,  422  _ 

See  also  alfalfa,  Bermuda,  clover,  cow-pea,  oat,  peanut,  timothy  and  vetch 
hay 
Heart,  cephalin  phosphorus — guinea  pig,  588 

composition,  affected  by  various  diseases,  507 

lecithin  content — children,  110,  111;  dog,  125,  443;  men,  125;  new-born 
infants  and  fetuses,  110,  111;  sheep,  125 
phosphorus — guinea  pig,  588 
mineral  analyses,  507 
phosphatid  content — guinea  pig,  588 
phosphorus — cattle,  126  . 
phosphocarnic  acid — dog,  124;  horse,  124 
phosphorus — guinea  pig,  588;  men,  507;  sheep,  149 
phosphorus  changes  effected  by  tuberculosis,  588 
Hedgehog,  lecithin  content,  107 
Helicoproteid,  occurrence,  47 
Hemaproton,  composition,  531 
Hemaprotagon  therapy,  531 
Heminucleic  acid,  definition,  20 

Hemiplegia,  effect  on  composition  of  spinal  cord,  538 
Hemorrhage,  influence  on  phosphorus  metabolism,  526 
Hemp  cake,  lecithin  (phosphatid)  content,  85 
seeds,  inorganic  phosphorus,  79 
lecithin  phosphorus,  79 
phosphorus,  79 
protein  phosphorus,  79 
Heparphosphatid,  composition,  69 
Herbivora,  carnivora  and  omnivora — acidosis,  478 

composition  of  blood,  369 
excretion  of  phosphorus,  207 
subcutaneous  administration  of  lactic  acid, 
553 
Histogehol,  description,  92,  93 
Hookworm,  phosphorus  metabolism,  527 
Hyperleucocytosis  after  nucleic  acids,  246,  247,  252 
Hypophosphites,  elimination,  206,  221,  222,  223,  226,  227 
feeding  experiments  with,  343-354 
influence  on  phosphates  in  intestine,  223 
in  urine,  222 

with  cod-liver  oil  in  treatment  for  rachitis,  576 
Hypophysis  and  thyroid  glands,  reciprocal  functions,  481 
effects  of  removal,  482 
phosphorus  content,  146 
studies  in  relation  to  acromegaly,  479 


INDEX  725 

Hypophysis  therapy,  diuretic  effect,  480 
in  acromegaly,  479 
with  hypoplastic  dwarf,  482 
Hypoxanthin,  composition,  18 

Hysteria,  phosphorus  content  of  cerebrospinal  fluid,  537 
Ichthulin,  chemistry,  47 
Icterus,  effect  on  phosphorus  metabolism,  527 

relative  phosphorus  and  chlorine  in  urine,  518 
Incubation,  phosphorus  metabolism,  446 
Indian  bunch  grass,  mineral  analyses,  affected  by  water  supplied,  94 

phosphorus,  affected  by  water  supplied,  94 
Infantile  athrepsia,  lecithin  therapy,  501,  529 
Infants'  bodies,  phosphorus  content,  108 
Infants,  phosphorus  metabolism,  449 

requirements,  414 
Inorganic  and  organic  phosphorus,  maintenance  experiments,  357 

metabolism  experiments,  320 
nutritive  value,  318,  355,  364,  427 
phosphates  and  nerve  stimulation,  463 

changes  in  egg  during  incubation,  447 
function,  in  muscular  action,  464 
metabolism  experiments  with,  221 
phosphorus  in  cancer  of  liver,  504 

salts,  absorption  of,  influenced  by  their  physical  condition,  376 
as  nerve  tonic,  546 

for  progressive  paralysis  and  for  psychoses,  545 
influence  on  amount  and  composition  of  milk,  382 
Inosinic  acid,  composition,  22 
Insanity,  effect  on  phosphorus  metabolism,  543 
Intestinal  extract,  digestion  of  nucleic  acids,  234 
infantilism,  phosphorus  absorption,  459 
juice,  digestion  of  nucleic  acids,  233,  245 
Iodocitin,  components,  533 
Iodothyrin,  distinction  from  thyroidin,  545 

effect  on  exophthalmic  goiter,  582 
Iodtriferrin,  metabolism,  247 

Iron  of  the  nucleoproteins,  15  * 

Jaundice.     See  Icterus 
Jecorin,  chemistry,  65 

Johnson  grass,  ether  extract  phosphorus,  90 
Kaffir  corn  fodder,  ether  extract  phosphorus,  90 
inorganic  phosphorus,  77 
mineral  analyses,  77 
phosphorus,  77 
Kidney,  cephalin  phosphorus — guinea  pigs,  588 

composition,  affected  by  various  diseases,  507 

diseases,  composition  of  blood  and  blood  serum,  498 

lecithin  content — cat,  107;  dog,  142;  hedgehog,  107;  rabbit,  107, 142 

phosphorus — guinea  pig,  588 
mineral  analyses — men,  507 
nuclein  content — cattle,  142 
partition  of  phosphorus,  142 
phosphatid  content — guinea  pig,  588 

effects  of  alcoholism,  484 
phosphocarnic  acid  content — dog,  142;  horse,  142 

phosphorus,  141;    cattle,  141,  142;     guinea  pigs,  588;    men,  141,  507; 
sheep,  149;  swine,  141,  373,  374 
content  and  distribution,  588 

in  tuberculosis,  588 
'Kufeke  mehl,"  calcium,  magnesium  and  phosphorus  balances — infants,  414 
Lactation,  effect  on  phosphorus  elimination,  209 
Lactic  acid  studies  in  osteomalacia,  552 


726  INDEX 

Lead  poisoning,  composition  of  blood  and  urine,  499 

Lecithid  in  rice,  80 

Lecithalbumins,  Liebermann's,  47,  66 

as  agents  in  urinary  excretion,  189 
Lecithin,  beet  leaves  and  roots,  87 
chemistry,  60 
cleavage,  asymmetric,  287 

by  colon  bacillus,  289 
Staphylococcus,  289 
succus  entericus,  289 
in  putrefaction,  537 
composition,  60 
compounds,  64 

dextro-  and  laevo-  forms,  287 
digestion  and  absorption,  processes,  286 

effects  on  composition  of  parts  of  the  body,  299,  300,  301,  343-354,  370 
ferment  activity  of  organs,  281 
ferratin  of  liver,  301 
growth,  296 

hydrochloric  acid  of  stomach,  551 
lipoid  deposits,  302 
milk  production,  301 

and  composition,  383 
phagocytic  index  of  leucocytes,  281 
power  of  oxidation,  280 
respiration  coefficient,  280 
weights  of  parts  of  body,  298,  299,  300,  301 
to  prolong  life,  298. 
feeding  and  injection  experiments,  280,  281,  286,  314,  341,  342,  343, 

343-354,  370,  383,  498 
function  as  antihaemolytic  agent,  280 

bringing  about  viscosity  of  cells,  278 
intermediary  product  in  oxidation  of  fats,  276,  280 
substratum  for  cell  reactions,  277 
in  digestion  of  fat,  173,  277,  282 
influence  on  diastase,  285 
,  in  intermediary  metabolism,  276 

relation  to  bone  formation,  568,  569 
significance  of  the  colloid  nature,  278 
in  relation  to  narcosis,  278,  280 
of  beet  leaves  and  roots,  87 
bile,  174 

in  relation  to  digestion  of  fat,  173,  277 
blood,  107 

as  affected  by  disease,  496,  511,  541,  542,  548 
of  different  species,  151,  152 
bodies  of  young  animals  in  relation  to  helplessness  at  birth,  277 
bone  marrow,  107,  118,  119,  542 

in  dementia  paralytica,    498,  542 
bones,  107,  506 

brain  and  nerves,  107,  130,  300,  539,  588 
as  affected  by  tuberculosis,  588 
of  human  fetuses  and  children,  110,  111 
eggs,  147,  171,  447 

during  incubation,  276,  341,  447 
ether  and  benzene  extracts  of  vegetable  substances,  86 
feces,  195,  196 

in  tabes  and  tabo-paralysis,  498 
sources,  288 
genitalia,  107,  147 
heart,  107,  125,  588 

as  affected  by  starvation,  443 
tuberculosis,  588 


INDEX  727 

Lecithin  of  heart,  in  health  and  disease,  125 

of  human  fetuses  and  children,  110,  111 
kidney,  107,  142,  588 

as  affected  by  tuberculosis,  588 
liver,  107,  137,  588 

as  affected  by  destruction  of  leucocytes,  494 

tuberculosis,  588 
in  relation  to  fatty  degeneration,  300,  513 

various  diseases,  513 
of  human  fetuses  and  children,  110,  111 
lungs,  588 

as  affected  by  tuberculosis,  588 
medulla,  588 

as  affected  by  tuberculosis,  588 
milk  at  different  periods,  166 

effect  of  pasteurization,  157 

identity  doubted,  158 

in  relation  to  brain  weight  of  the  young,  277 

the  buttermilk,  157 
of  different  species,  155,  162,  169,  450 
skimmed  milk  and  cream,  163 
miscellaneous  common  foods,  87 
muscles,  107,  122,  125,  588 

as  affected  by  nerve  stimulation,  463 
starvation,  443 
tuberculosis,  588 
of  human  fetuses  and  children,  110,  111 
nitrogenous  concentrates,  85,  91 
pancreas,  141 
pancreatic  juice,  141 
parts  of  the  body — rabbits,  107 
plants  and  in  miscellaneous  foods,  84 
sperm,  148 

spinal  cord  after  hemiplegia,  538 
spleen,  140,  588 

as  affected  by  tuberculosis,  588 
suprarenal  capsules,  143 
thymus,  143 
phosphorus  of  milk,  156,  157 

of  different  species,  157 
seeds,  grains  and  cereal  products,  79,  80,  82,  85,  87,  89 
properties,  63,  284  »»_.#.*» 

synthesis  in  developing  eggs,  341 
egg  production,  342 
therapy,  528 

after  activation  by  Rontgen  rays,  532 
effect  on  narcosis,  546 
in  anaemia,  501 

exophthalmic  goiter,  582 
lues,  tabes,  syphilis  and  paralysis,  497,  546 
migraine,  547 
neurasthenia,  545 
tabes,  546 
_     .  ,  tuberculosis,  585 

Lecitho-proteins,  66 
Lecitogen,  lecithin  content,  501 

treatment   in  anaemia,  501 
Lentil  seeds,  inorganic  phosphorus,  79 

lecithin  (phosphatid)  content,  85 

phosphorus,  79,  81 
phosphorus,  79,  81,  85 
phytin  phosphorus,  81 
protein  phosphorus,  79 


728  INDEX 

Lettuce,  phosphatid  phosphorus,  89 
Leucocytosis  in  blood  dissolution,  496 

induced  by  bromlecithin,  501 

lecithin  ingestion,  533 
injection,  530 
nucleate  ingestion,  519 
injection,  547 
nucleic  acid  injection,  258 
nucleins,  247,  257,  502 
pneumonia,  499 
phytin,  533 
See  also  Leukaemia 
Leukaemia,  effects  on  composition  of  blood,  497,  498 

phosphorus  metabolism,  493 
Linseed  oil  meal  and  corn,  specific  effects  on  swine,  376 
inorganic  phosphorus,  78 
mineral  analyses,  78 
phosphorus,  78 
Lipanin  treatment  in  rachitis,  567,  573 
Lipoid  content  of  brain  as  affected  by  age— dog,  135 
organs,  588 

as  affected  by  tuberculosis,  588 
phosphorus  estimations  in  nerve  tissue,  134 

spleen,  139 
Lipoids,  absorption  by,  280 
definition,  58 

essential  for  life,  358,  362,  363 
Liver,  cephalin  phosphorus — guinea  pig,  588 

composition  as  affected  by  cancer,  497,  504 

various  diseases,  507 
ether-alcohol  soluble  phosphorus — rabbit,  313 
ferratin,  effects  of  organic  phosphorus  compounds  on,  301 
lecithin  content — cat,  107,  137;  children,  110,  111;  hedgehog,  107;  new- 
born infants  and  fetuses,  110,  111;  rabbit,  107,  137 
as  influenced  by  alcohol  poisoning,  484 

destruction  of  leucocytes,  494 
fast,  446 

various  diseases,  513 
in  fatty  degeneration,  513 
phosphorus — guinea  pig,  588 
mineral  analyses — men,  105,  507;  rabbit,  313;  swine,  327 
nuclein  phosphorus — cattle,  137;  dog,  137;  fowl,  137;  new-born  infants 

and  fetuses,  138;  rabbits,  138 
nucleoprotein  content — rabbit,  139 
pathological  changes  of  the  phosphorus  content,  137 
phosphatid  content — guinea  pig,  588 
phosphatids  of,  differentiated,  138 
phosphocarnic  acid— dog,  138;  horse,  138 

phosphorus — cattle,  107,  136,  137,  149;  dog,  137;  fowl,  137;  guinea  pigs, 
588;  men,  104,  105,  507;  new-born  infants  and  fetuses,  136; 
rabbits,  313;  sheep,  136;  swine,  137,  327,  373,  378 
relation  to  nuclein  metabolism  studied  by  means  of  Eck  fistula,  240 
Livetin,  occurrence,  45 
Lues,  spinal,  lecithin  content  of  blood  serum,  500 

lecithin  therapy,  531,  546 
Lung  diseases,  phosphaturia  in.     See  Phosphaturia 
Lungs,  cephalin  phosphorus — guinea  pig,  588 

composition,  as  affected  by  pneumonia,  507 

lecithin  content — cat,  107;  hedgehog,  107;  rabbit,  107 

phosphorus — guinea  pig,  588 
mineral  analyses — men,  105,  507 


INDEX  729 

Lungs,  phosphatid  content — guinea  pig,  588 

as  affected  by  alcoholism,  484 
phosphorus — guinea  pigs,  ^588;  horse,  146;  men,  104, 105, 507;  sheep,  149 
compounds,  145 
Lymph,  mineral  analyses — men,  105 

phosphorus — horse,  177;  men,  105,  177 
Magnesium,  substitution  for  calcium  in  bone,  386,  390 
Maintenance  of  neutrality,  178,  181 
Malnutrition  of  bones,  534 

called  pseudorachitis,  573 
contrasted  with  osteomalacia,  552 
Malt,  lecithin  (phosphatid)  content,  89 

phosphorus,  89 
Mange,  benefits  from  nucleins,  257 
Mangel  wurzel,  inorganic  phosphorus,  78 
mineral  analyses,  78 
phosphorus,  78 
Massage,  effect  on  phosphorus  excretion,  466 
Meadow  fescue,  phosphorus  as  affected  by  water  supplied,  93 
Meat,  ash,  effects  on  growth,  227 

inorganic  phosphorus,  78,  120 

lecithin  content,  87,  104,  107,  125 

light  and  dark  compared  as  to  purin  excretion,  525 

meal  added  to  corn,  specific  effects  on  swine,  376 

mineral  analyses,  78,  121 

nuclein  content,  104 

and  phosphoprotein  phosphorus,  88 
phosphatid  phosphorus  content,  88 
phosphocarnic  acid  content,  104 
phosphorus,  78,  88,  104,  107,  120,  121 
ration  compared  with  edestin,  338,  339 
Meconium,  mineral  analyses,  194 

phosphorus,  194 
Medulla,  organic  phosphorus  compounds — guinea  pigs,  588 

phosphorus — guinea  pigs,  588 
Melancholia,  lecithin  therapy,  531 
Mellin's  Food,  analysis,  451 

as  a  milk  supplement  for  infants,  451 
compared  with  human  milk  in  composition,  451 
Mental  and  other  nervous  disorders,  effects  on  phosphorus  metabolism,  536 

use  of   drugs  and   effects   on  phosphorus 
metabolism,  545 
Mental  work,  influence  on  phosphorus  metabolism,  461 
Metabolism  in  osteomalacia,  556 
of  casein,  260 

general  summary,  274 
glycerophosphoric  acid  compounds,  276 
inorganic  phosphates,  221,  318 
nucleoproteins  and  nucleic  acids,  229 
phosphocarnic  acid,  303 

phosphorus  in  relation  to  other  elements,  213 
phytin,  305 
See  also  Digestion 
Metacasein  formation  by  pancreatic  extract,  269 
Metaphosphates,  poisoning  from,  226 
Metaphosphoric  acid,  poisonous  effects,  221 

poisoning  from,  227 
Migraine,  lecithin  treatment,  547 

Milk,  calcium  content,  as  affected  by  calcium  salts,  571,  573 
range  of  variation,  158 
casein  content,  160;  ass,  155,  161;  cow,  155,  161,  163,  164,  166;  goat,  161; 
others,  161;  woman,  155,  161, 167 


730  INDEX 

Milk,  casein  content,  affected  by  tuberculosis,  382 

of  human,  factors  influencing,  162 
phosphorus — ass,  156;  cow,  156,  157;  woman,  156,  157 
changes  resulting  from  heating  or  standing,  164 

with  progress  of  lactation,  165 
component  essential  to  normal  growth,  358,  360,  362 
composition,  compared  with  that  of  the  young,  167 

in  relation  to  time  required  for  young  to  double  in  weight, 
168 
fermented,  organic  phosphorus  compounds,  162 
glycerophosphoric  acid  phosphorus — cow,  163,  166 
inorganic  phosphorus — ass,  156;  cow,  154,  156,  157,  165 
lecithin  content— ass  155,  162;  cow,  87,  155,  162,  163,  166,  169,  450;  goat, 
162;  others,  162,  169;  woman,  87,  155,  162,  169,  450 
phosphorus— ass,  156;  cow,  156,  157,  163, 166,  450;  woman,  156, 157 
metabolism  experiments,  273,  274,  322,  326,  367,  414,  417,  449,  456,  457, 

518,  570 
mineral  analyses— ass,  159;  cow,  159;  goat,  159;  others,  159;  woman,  105, 

159,  160,  418 
modification  of  cow's,  for  human  infants,  417,  449,  450 
nuclein  and  phosphoprotein  phosphorus — cow,  88 

nucleon  content-ass,  155,  163;  cow,  155,  162,  163;  goat,  163;  woman,  155, 
163 
phosphorus — goat,  157;  cow,  157;  woman,  157 
of  different  species  compared  in  diet  of  man,  367 

infant  feeding,  449 
phosphatid  content,  162 

as  affected  by  pasteurization,  157 
doubted,  158 

in  the  brain  weight  of  young  animals,  277 
phosphorus  content — cow,  88,  162 
phosphocarnic  acid  (nucleon)  content,  162 

phosphorus— ass,  156,  159;  cow,  88,  156,  157,  159,  160,  163,  164,  165,  166, 
168,  380,  381,  450;  goat,  157,  159,  168,  383;  others,  159, 
168;     woman,     105,  156,  157,  159,  160,  167,  168,  405, 
418,   450 
content     as     affected    by     inflammation     of  the  udder  and 
tuberculosis,  382 
protein  phosphorus — woman,  158 
salts  of,  160 
secretion  as  affected  by  foods,  379 

lecithin,  301 
phytin,  307,  312,  315 
effect  on  phosphorus  requirement,  389 
utilization  of  minerals  from,  456,  457 
whey  feeding  for  infants,  455 
See  also  Skimmed  milk 
Millet  bran,  lecithin  phosphorus,  82 
phosphorus,  82 
hay,  ether  extract  phosphorus,  90 
inorganic  phosphorus,  78 
mineral  analyses,  78 
phosphorus,  78 
Mineral  constituents  of  blood  under  normal  conditions,  501 

human  organs  after  death  from  various  diseases,  507 
milk,  158 
elements  in  the  urine  during  fast,  400 
matter  of  bones     as  obtained     by  removal     of  organic     matter  with 

glycerin  and  potassium  hydrate,  113 
retention  by  the  fetus,  471 
Monamino-diphosphatids,  68 

-monophosphatids,  60 


INDEX  731 

Mongoloid  idiocy,  thyroid  treatment,  546 
Mononucleic  acids,  definition,  21 
chemistry,  21 
Mononucleotides,  definition,  21 

Morbus  basedowii  in  relation  to  phosphorus  metabolism,  581 
Muscle  phosphatids  differentiated,  125,  126  , 

Muscles,  comparisons  of  striated  and  non-striated,  123 

cephalin  phosphorus— guinea  pig,  588  ' 

inorganic  phosphorus— cattle,  120,  126;  dogs,  468;  rabbit,  463;  sheep,  78 
lecithin  content— children,  110,  111;     dog,  125,  443;    hedgehog,  107; 
new-born  infants  and  fetuses,  110,  111;  rabbit,  107,  463; 
sheep,  125 
phosphorus — fish,  122;  guinea  pig,  588 
mineral  analyses— cat,  121;  cattle,  121;  deer,  121;  dog,  121;  fish,  121; 
fowl,  121;  frog,  121,  124;  men,  105,  121;  rabbit,  121;  sheep,  78; 
swine,  121,  124,  327 
nuclein  and  pseudonuclein  phosphorus — fish,  122,  147 

phosphorus — cattle,  122;  fowl,  122 
phosphatid  content — guinea  pig,  588 

phosphorus — cattle,  126 
phosphocarnic  acid — dog,  305;  men,  123;  new-born  infants  and  fetuses, 
123;  rabbit,  304 
phosphorus — dog,  468 
phosphorus— cat,  121;  cattle,  107,  120,  121,  122,  123,  126,  149;  children, 
458,  572;  deer,  121;  dog,    121,  468;     fish,  121,  122,  146-147; 
fowl,  121,  122;  frog,  121,  122,  124 
phosphorus— guinea  pig,  588;     men,  104,  105,  121,  462;  rabbit,  121; 

sheep,  78;  swine,  121,  124,  327,  370,  372,  373 
phosphorized  proteins  spared  during  atrophy,  445 
phosphorus  changes  effected  by  exercise — dogs,  468 

nerve  stimulation,  463 
tuberculosis,  588 
compounds,  120 
See  also  Heart 
Muscular  action,  in  relation  to  lactic  acid  and  potassium  phosphate,  464  _ 

formation  and  phosphorus  elimina- 
tion, 465 
Mushrooms,  lecithin  (phosphatid)  content,  85,  87 
Myosin,  feeding  experiment,  320 
Myostromin  formation  from  myosin,  293 
Myxoedema  attributed  to  hypophysis  and  thyroid  gland,  481 
thyroid  treatment,  546,  578 

vegetable  diet  and  disodium  phosphate  treatment,  546 
Neottin,  chemistry,  71 
Nephritis,  effects  on  blood,  498,  500,  548 

and  urine,  499 
phosphorus  metabolism,  547 
purin  excretion  delayed,  525 
Nerve  degeneration,  changes  in  phosphorus  compounds,  536 

katabolism,  nitrogen  and  phosphorus  ratio,  216 
Nerves,  composition.    See  Brain  and  Nerves 
Nerve  stimulation,  electrical,  phosphoric  acid  change,  463 

influence  on  phosphorus  metabolism,  463 
Nervous  disorders,  effects  of  drugs  on  phosphorus  metabolism,  545 

on  phosphorus  metabolism,  536 
disturbances  as  cause  of  phosphaturia.     See  Phosphaturia 
excitement,  effect  on  urinary  phosphorus,  462 
Nervinol,  description,  93 

Neuralgia,  chronic,  glycerophosphate  treatment,  521 
Neurasthenia,  glycerophosphate  treatment,  521 
lecithin  treatment,  545,  585 
phosphorus  content  of  cerebrospinal  fluid,  537 

excretion  under  excessive  protein  feeding,  545 


732  INDEX 

Nitrogen  and  phosphorus  ratio  in  feces  during  pregnancy,  472 

fever,  explained,  519 
muscle,  influence  of  degeneration,  445 
organs,  influence  of  fast,  445 
urine,  influenced  by  cow's  and  human  milk, 
451,  456;   epilepsy,  541;   exercise,  469; 
experimental  diphtheria  infection,  516; 
fast,  401,  402,  438,  439,  440,  442,  443, 
644;  fever,  514,  516;  hypophysis  treat- 
ment, 481;  lmyphatic  leukaemia,  495;  a 
meat  diet,  215;  nephritis,  548;  phases  of 
malarial  fever,  516;     pregnancy,     472; 
sleep,  472;     state     of    nutrition,     215; 
sweating,  467;  thyroidectomy,  580;  vari- 
ous diseases,  563 
See  also  Phosphaturia 
Normal  phosphorus  metabolism,  178 

Nuclease  content  of  blood  serum  in  nervous  diseases,  540 
organs,  effect  on  thyroidectomy,  580 
definition,  28 

influenced  by  administration  of    nucleic  acids,  253,  254 
in  the  stomach,  age  at  which  it  appears,  233 
occurrence,  235 
Nucleic  acid  cleavage  by  organ  ferments,  235 
enzymes  concerned,  28 
partial,  20 
possible  paths,  29 
acids,  absorption,  244 

a  and  ft,  definition,  20 
"biological  value,"  254 
chemistry,  16 
digestion  by  erepsin,  232 
peptic,  231 
tryptic,  231 
effects,  miscellaneous,  250 

on  action  of  Bacillus  coli,  252 

Staphylococcus  infection,  250 
composition  of  parts  of  body,  301 
enzyme  activities  of  the  organs,  250 
flow  and  composition  of  milk,  383 
hydrochloric  acid  of  the  stomach,  551 
leucocytosis,  248,  252 
power  of  oxidation,  280 
purin  excretion,  245 
respiration  coefficient,  280 
tubercle  bacilli,  257 
weights  of  parts  of  the  body,  301 
feeding  and  injection  experiments,  237,  247,  343-347 

See    also    Nucleic    acids,    effects, 
and  Nucleic    acids,  therapy    and 
prophylactic  use 
functions  as  active  agents  in  specific  body  processes,  252,  255 
in  fish  eggs,  171 
intestine,  146 
metabolism,  229 

methods  of  introduction,  comparison  of  results,  249 
prophylactic  use,  258,  504 
therapeutic  and  prophylactic  use,  256 
Nucleic  acid  therapy,  abscesses,  502 
paralysis,  547 
scarlet  fever,  519 
tuberculosis,  with  disodium-methyl-arsenate,  586 


INDEX  733 

Nuclein,  definition,  13  _ 

-free  diet  in  gout,  nitrogen  and  phosphorus  balances — men,  524 
in  blood,  151,  152;  bones,  506;  brain  and  nerves,  130,  134;  cereal 
products,  79,  88;  eggs,  88;  fetuses,  138;  genitalia,  147;  kidney,  142; 
liver,  137,  138;  meat,  88,  104;  milk,  88;  muscle,  122,  147;  nitrogen- 
ous concentrates,  90,  91;  pancreas,  141;  rice,  80,  88;  seeds,  79; 
spleen,  139,  140 

suprarenal  capsules,  143;  thymus,  143;  vegetables,  88 
katabolism  in  exercise  reduced  by  training,  466 
metabolism,  229 

in  gout,  523 
phosphorus  distribution  in  foods,  90 
synthesis,  30,  229,  341,  354 
therapy,  256 

in  anaemia,  502 
chlorosis,  502 
Nucleins,  digestion,  239 

ferments  of  individual  organs,  235 
peptic,  231 
tryptic,  231 
effects  on  bacteria,  252 

composition  of  parts  of  the  body,  301 
flow  and  composition  of  milk,  383 
leucocytosis,  247 

weights  of  parts  of  the  body,  301 
feeding  experiments,  238,  301,  309,  343,  383,  522 
Liebermann's,  feeding  experiments,  322 
prophylactic  use,  252 
therapeutic  and  prophylactic  use,  256 
Nucleinases,  definition,  30,  236 

occurrence,  236 
Nucleoalbumins  of  eggs,  170 
Nucleohistone,  definition,  144 

occurrence,  144 
Nucleon.     See  Phosphocarnic  acid 
Nucleoprotein  digestion,  accompaniments,  245 

direct  observations  on  the  alimentary  tract,  244 
effects  of  bacteria  in  the  alimentary  tract,  234 
peptic,  232 
tryptic,  231 
katabolism,  methods  of  experimental  study,  232 
Nucleoproteins,  chemistry,  13 

cleavage  products,  17 
composition,  15 

feeding  experiments  with,  237,  322 

formed  from  phosphoproteins  during  incubation,  446,  447 
functions  as  active  agents  in  specific  body  processes,  255 
in  bile,  174;  brain,  nerve,  spinal  cord,  130;  eggs,  147;  genitalia, 
147;  liver,  139;  pancreas,  141;  pancreatic  juice,  141,  173; 
spleen,  140;  thymus,  143;  thyroid,  144 
metabolism,  229 
rectal  feeding,  239 
Nucleosidases,  definition,  30,  236 

occurrence,  236 
Nucleosides,  definition,  21 
Nucleothyminic  acid,  definition,  21 
Nucleotidases,  definition,  30,  236 

occurrence,  236 
Nucleotides,  digestion,  233,  236 
Nucleotin,  definition,  21 
Nutrose,  composition,  92,  237,  320 

feeding  experiments,  237,  320 


734  INDEX 

Oat  grains,  inorganic  phosphorus,  77 
mineral  analyses,  77,  422 
phosphatid  phosphorus,  89 
phosphorus,  77,  83,  89,  96,  97,  422 
hay,  ether  extract  phosphorus,  90 
hull,  phosphorus,  83 
kernel,  phosphorus,  83    ' 

plants,  phosphorus  as  affected  by  water  supplied,  93,  94 
Oats  compared  with  hay  in  effect  on  bones,  389 

phosphorus  compounds,  as  affected  by  composition  of  soil,  96 
phytin,  53 
Oedema,  effect  of  varying  water,  sodium  chloride  and  phosphorus  content  of 

the  food,  549 
Oil  cakes,  flax,  lecithin  (phosphatid)  content,  85 
Onions,  inorganic  phosphorus,  78 
mineral  analyses,  78 
phosphorus,  78 
Oophorin  treatment  for  osteomalacia,  558 

Organic  and  inorganic  phosphorus,  comparative  nutritive  value,  summary,  364 
phosphorus  compounds,  chemistry,  13 
in  the  urine,  190 

changes  in  lymphatic  leukaemia,  495 
low  requirement,  330 
Organs,  composition  as  affected  by  disease,  507 
Orizanine  an  essential  nutrient,  490 
cures  polyneuritis,  490,  491 
occurrence,  491 
properties,  490 
Ortho-,  meta-  and  pyrophosphates,  comparison  in  animal  tests,  226 
pyrophosphoric  acids,  chemical  comparison,  227 
pyro-  and  metaphosphoric  acids  as  anticoagulating  agents,  256 
Osteomalacia,  552 

distinct  from  malnutrition  of  the  bones,  halisteresis  of  the  bones, 

fragility  of  the  bones  and  osseous  cachexia,  552 
effects  on  blood,  557 

bone  formation  attributed  to  lecithin,  569 
composition  of  blood  ash,  557 

bones,  554 
parathyroids,  559 
etiology  as  a  phosphorus  metabolism  disturbance,  485 
due  to  excessive  lactic  acid  production,  553 
disease  of  the  thyroid  gland,  558 
experimental,  due  to  low  calcium  diet,  394     . . 

effects  of  dicalcium  phosphate  and  of  bone,  394 
metabolism,  556 

of  calcium,  magnesium  and  phosphorus,  559 
methods  of  treatment,  558 
Ovarian  preparations,  effects  on  metabolism  of  spayed  and  normal  dogs,  506 
Ovariotomy,  influence  on  metabolism  and  composition  of  the  body,  506 
in  osteomalacia,  558 

influence  on  calcium,  magnesium  and  phosphorus 
metabolism,  558,  559 
Ovovitellin,  chemistry,  45 

maintenance  experiments,  360 
tryptic  digestion,  271 
Oysters,  phosphocarnic  acid  content,  123 
Palm  cake,  lecithin  (phosphatid)  content,  85 

nuclein  phosphorus,  90 
Pancreas,  in  relation  to  diabetes,  510,  511 

organic  phosphorus  compounds — cattle,  141;  dog,  141 
partition  of  phosphorus,  141 

phosphorus  content,  140;  cattle,  141;  children,  141;  men,  141;  new- 
born infants,  141;  sheep,  149 


INDEX  735 

Pancreas,  removal  of,  effect  of  calcium,  nitrogen  and  phosphorus  of  the  urine 

of  a  fasting  dog,  512 
Pancreatic  digestion  of  casein,  in  vitro  tests,  267 

fats,  relation  of  bile  to,  282 

lecithin  to,  282 
extract,  digestion  of  nucleic  acids,  234 
juice,  digestion  of  nucleic  acids,  233,  245 
nuclein,  233 
inorganic  phosphorus — dog,  173 
organic  phosphorus  compounds — dog,  141,  172 
phosphorus,  172;  cattle,  173;  dog,  173 
Papain  digestion  of  casein,  271 
Parachymosin,  occurrence  and  action,  38 
Paralysis  agitans,  thyreoidin  therapy,  545 

and  senility,  thyreoidin  therapy,  545 
effects  on  phosphorus  metabolism,  541 
general,  changes  in  the  brain,  540 
lecithin  treatment,  546 

progressive,  effect  on  ferment  activity  of  cerebrospinal  fluid,  541 
lecithin  content  of  serum,  500 
phosphorus  content  of  cerebrospinal  fluid,  537 
treatment  with  inorganic  salts,  545 
nucleic  acid,  547 
nucleins,  257 
Paranucleic  acids  from  phosphoproteins,  33 
Paranucleins,  definition,  14 

from  phosphoproteins,  33 
Parathyroid  treatment,  effect  on  bone  formation,  559 
Parathyroidectomy,  investigations  of  the  results,  559 
Parathyroids  in  relation  to  formation  of  dentine  and  enamel,  560 

osteomalacia,  559 
phosphorus  metabolism,  559 
Peanut  cake,  lecithin  (phosphatid)  content,  85 
nuclein  phosphorus,  90 
hay,  ether  extract  phosphorus,  90 
Peanuts,  inorganic  phosphorus,  78 
mineral  analyses,  78 
phosphorus,  78 
Peas,  lecithin  content,  87 

(phosphatid)  content,  85 
phosphorus,  81 
phosphorus,  81,  85 
phytin  phosphorus,  81 
(yellow),  inorganic  phosphorus,  79 

nuclein  and  phosphoprotein  phosphorus,  79 
phosphatid  phosphorus,  79 
phosphorus,  79 
phytin  phosphorus,  79 
Peptic  digestion  of  nucleoproteins,  232 
casein,  262 

affected  by  carbohydrates,  267 
as  a  reversible  action,  266 

comparison  of  casein  from  different  species,  265,  267 
phosphorus  of  the  nuclein  resulting  from,  263 
results,  264 

Salkowski's  description  of  the  process,  264 
under  different  conditions  of  acidity,  268 
Phosphate  of  calcium,  amounts  to  be  used  for  different  animals,  535 
deposits  under  the  skin,  503 
for  diabetes  mellitus,  511 

experimental  osteomalacia,  394 
influence  on  composition  of  parts  of  body,  327,  328,  385, 


736  INDEX 

Phosphate  of  calcium  with  phosphorized  cod-liver  oil,  for  rickets,  576 

influence  on  composition  of  parts  of  body,  327,  328,  385, 
386,  388,  389 
milk  composition  and  production,  379,  380, 
383 
other  feeding  experiments,  222, 223,  322, 324, 326, 828 
precipitated,  compared  with  bone  ash,  floats  and  bran 

in  feeding  experiments,  328 
See  also  Bone  Meal 
potassium,  feeding  experiment,  324 

produces  a  rachitic  effect  in  bones,  569 
sodium  compared  with  organic  phosphorus  compounds  in  feeding 
experiments,  312,  314,  326,  335,  343 
for  cretinism,  579 

myxoedema  with  a  vegetable  diet,  546 
influence  on  composition  and  production  of  milk,  381,  383 
of  parts  of  body,  313,  370,  385, 
389,  391 
injection,  influence  on  excretion  as  affected  by  species,  209 
Phosphates,    absorption,  182 

acid  sodium  phosphate  unfavorable  to  the  use  of  oats,  389 
addition  to  silage,  101 

and  lecithin  content  of  muscle  as  affected  by  nerve  stimulation,  463 
effect  on  composition  of  bones,  385 

milk,  380 
effects  on  growth,  228 

nausea  and  catharsis,  357 
feeding  experiments,  342 

See  also  the  individual  phosphates 
influence  on  digestion,  186 

enzyme  action,  176,  186 
fermentation,  31 
in  maintenance  of  neutrality,  178 
intestinal  changes,  184 
method  of  elimination  in  the  urine,  188 
of  alkaline  earths,  feeding  experiments,  221,  222,  224,  225 
bones,  effects  of  diet,  384 
sodium  and  calcium,  feeding  experiments,  326 

potassium  compared  with  lecithin  and  with  a  mix- 
ture of  phosphorus  compounds  for  maintenance 
of  life,  343 
urine  as  affected  by  acid  in  the  blood,  445 

pneumonia  and  typhoid  fever,  519 

time  of  day    and  increased    ingestion  of 

protein,  475 
various  diseases,  561 
paths  of  elimination,  182 
Phosphate  therapy  in  psychasthenia,  545 
psychatoxies,  545 
See  also  the  individual  phosphates 
Phosphatese,  definition,  32 

essential  in  organic  phosphorus  synthesis,  491 
in  cure  of  polyneuritis  and  beriberi,  491 
Phosphatid  content  of  blood  as  affected  by  anaemia,  496 

chlorosis,  496 
eggs,  171 
liver  as  affected  by  cancer,  504 

of  incubating  chick,  449 
milk,  162 
organs  as  affected  by  alcoholism,  484 

tubercle  bacilli,  dead,  587 
tuberculosis,  587 
definition,  58 
deposits  as  a  result  of  lecithin  and  egg  yolk  feeding,  302 


INDEX  737 

Phosphatid,  synthesis,  74,  341 
Phosphatids,  chemistry,  58 

classification  by  Bang,  59 

Rosenheim,  58 
differentiation  in  blood  cells,  153;  egg  yolk,  171;  flours,  89;  heart 
muscle,  125,  126;    kidney,  142;     liver,  138;     lungs,  145; 
spleen,  140;  suprarenal  capsules,  143 
function  as  semipermeable  membrane,  278 

in  brain,  135,  588;  heart,  588;  kidneys,  588;  liver,  588;  lungs,  588; 
medulla,  588;  muscles,  588;  spleen,  588;  suprarenal  capsules,  143 
naming  of,  58 
"plant,  chemistry,  73 

physico-chemical  reactions  with  inorganic  and  organic  substances, 
278 
Phosphaturia,  561 

following  quinine  treatment  for  fever,  515 
in  artificially  fed  infants,  459 

tuberculosis,  effect  of  arsenic-phosphorus  treatment,  586 
phytin  treatment,  315 
Phosphocarnic  acid,  chemistry,  49 

cleavage  products,  49 

function  as  a  carrier  of    mineral    elements    in  the  body 
fluids,  303 
an  energy-producing  nutrient,  304 
a  source  of  muscular  energy,  467 
in  blood,  150;  brain  and  nerves,  130,  305;  genitalia,  147; 
kidnev,  142;  liver,  138;  meat,  104;  milk,  155,  157,  162, 
163;     muscle,     123,  124,  304,  305,  468;     spleen,     140; 
urine,  191 
brain,  effect  of  fast,  305 

nervous  excitement,  305 
muscle,  factors  influencing  amount,  303 
metabolism,  303 
Phospholipins  in  genitalia,  148 
Phosphoprotein,  definition,  33 
Phosphoproteins,  chemistry,  32 

in  eggs,  147;  genitalia,  147;  pancreas,  141;  pancreatic  juice, 

141,  173 
occurrence,  147 
of  animal  origin,  47 
vegetable  origin,  48 
Phosphoric  acid  administration,  benefits  from,  224 

in  dyspepsia,  223 
in  case  of  hypochlorhydria,  223 
results  in  rheumatism,  224 
Phosphorized  proteins  spared  in  degenerating  muscle,  188 

fever,  519 
Phosphorus  absorption  during  rectal  feeding,  404 

in  intestinal  infantilism,  459 
effect  of  low  fat  content  of  diet,  568 
and  calcium  balances  with  cattle,  222;  dogs,  225,  323,  435;  infants, 
414,  416,  417,  418,  453,  455,  570;  growing  children,  411,  412, 
413,  453;  men,  218,  219,  293,  326,  403,  408-409,  512;  sheep, 
221;  swine,  375 
phosphatid  content  of  organs  as  influenced  by  chronic  tubercu- 
losis, 588 
vaccination    with 
dead  tubercle  ba- 
cilli, 587 
balances  with  calves,  diet  of  skimmed  milk  and  mineral  supple- 
ments, 222 


738  INDEX 

Phosphorus  balances  with  children  convalescent  from  scarlet  fever  on  a  diet 

of  milk,  518 
cows,  high  and  low  intake  compared,  420 

phosphorus  rations  compared,  420 
rations  of  different  common  foods,  420 
dogs  as  affected  by  external  hemorrhage,  526 
inorganic  salts,  225 
phytin,  311 
diets  varying  in  amount  of  fat,  carbohydrate 

or  protein,  435 
increasing  amounts  of  food,  437 
normal  rations  supplemented  by  inorganic  salts, 

205 
organic  and  inorganic  phosphorus  compared,  323 
growing  boys,  ordinary  mixed  diet,  511,  512 

taking  different  amounts  of  exercise, 
413 
horse,  low  calcium  -rations,  423 
infants,  artificially  fed,  414,  416 
breast-fed,  418 

diet  of  milk  gruel  and  malt,  455 
healthy^    and     diseased,     on     natural     and 

artificial  milk  diet,  452 
milk  diets  of  varying  composition,  417 
normal  and  rachitic,  on  milk  diets,  570 
organic     and     inorganic    phosphorus    com- 
pounds compared,  325 
raw  and  sterilized  milk,  computed  to  kilo- 
gram body  weight,  416,  453 
men,  218 

as  influenced  by  antirabes  with  and  without  sodium 
glycerophosphate,  521 
casein  added  to     normal     mixed 

diet,  293 
excessive  water  drinking,  473 
fatigue   of  mountain  climbing,  469 
lecithin,  291 

added  to  mixed  diet,  295 
loss  of  sleep;     crackers,     butter, 

_  and  milk  diet,  432 
nuclein-free  diet  in  gout,  524 
nucleins  added  to  mixed  diet,  238 
nucleins,   gout  patient   and  con- 
trol, 522 
phosphate  of     sodium     added  to 

ordinary  mixed  diet,  202 
phytin  added  to  simple  diet,  486 

overfed  condition,  306 
sanatogen  added  to  normal  mixed 
diet,  293 
at  different  altitudes,  429 

diet  low  in  phosphorus  with  supplements  added 
at  intervals,  407 
of  crackers  and  milk,  409 
diets  composed  principally  of  different  sorts  of 
rice,  488 
containing  different  amounts  of  phosphorus, 

.486.  ' 

varying  in  phosphorus  and  nitrogen    con- 
tent, 433 
content,  404 
protein  content,  433 


INDEX  739 

Phosphorus  balances  with  men,  endoarteriitis,  512 

following  a  fast,  402 
normal  mixed  diet,  403,  406 
overfeeding,  219 

phosphate,  glycerophosphate  and  phytate  of  cal- 
cium compared  as  added  to  mixed 
diet,  324 
of  calcium  and  casein  (proton)   com- 
pared as  added  to 
bread     and     milk 
diet,  322 
and  sodium  compared  with 
milk  in    simple     diet, 
326 
senility  and  paralysis   agitans     as  affected  by 

thyreoidin,  545 
simple  mixed  diet,  411 

whole  wheat  bread  and  white  bread  compared, 
366 
rabbits,  phytin  and  sodium  phosphates  compared,  312 
rats  as  influenced  by  phosphorus  added  to  nitrogen- 
free  rations,  340 
powdered  beef  and  edestin  compared,  338,  339 
without  organic  phosphorus,  335 

with    and    without 
protein,  337 
sheep  as  influenced    by  phosphates    of  calcium  and 
magnesium  added     to  a  ration  of  meadow 
hay,  221 
normal  foods,  426 
swine  as  influenced  by  nucleic  acids,  242 

cereal  foods  with     and     without     additional 

calcium,  375 
common  food  mixtures,  378 
different  types  of  phosphorus  compounds,  328 
young,  424 
women  during  progress  of  pregnancy,  472 
See  also  Phosphorus  Requirements 
calcium  and  magnesium  metabolism  as  influenced  by  ovariotomy  in 

case  of  osteomalacia,  558,  559 
complexes    artificially    prepared,    investigations    of   physiological 

effects,  549 
content  of  dietaries,  409,  410 

foods,  references,  103 

organs,  urine  and  feces  in  relation  to  nitrogen  content. 

See  Nitrogen  and  phosphorus  ratio 
young  dogs  as  influenced  by  removal  of  certain  organs, 
580 
distribution  in  different  cuts  of  beef,  120 

muscles  as  affected  by  exercise — dogs,  468 
elemental,  effects  on  mineral  metabolism  in  dogs,  577 
excretion  as  influenced  by  acids,  salts,  diet  and  species  of  subject, 

198 
exercise,  sweating  and  massage,  466 
fast,    399,  426,  466;   fat,  459;     mental 
work,  461;  oil,  576;     rachitis,  573, 
574;   sodium  carbonate  in  acidosis, 
510;  sweating,  467;  wine  drinking, 
469 
in  fever,  variations  explained,  519 

relation  to  chlorine  in  various  diseases,  518 


740  INDEX 

Phosphorus  elimination  in  relation  to  urea  and  to  sulphur,  effects  of  exercise, 
464 
loss  during  fast;  sources  of  the  phosphorus,  438 
metabolism  as  influenced  by  alcohol  ingestion,  483;  altitude,  429; 
amount  of  food,  430,  458;  castration  and  ovarioto- 
my, 505,  559;    exercise,   464;       hemorrhage,   526; 
hypophysis  therapy,  479;  nerve  degeneration,  536; 
nerve  stimulation,  463;  parathyroids,  559;   thirst 
and  water  drinking,  473;  thyroid  glands,  577;  time 
of  day,  474;  toxins  and  antitoxins,  582 
during  fast,  438;  fever,  514;  first  few  days  of  infant 
life,  414,  458;  hypnotic  sleep,  442;  incubation,  446; 
infancy,  449;  pregnancy,  470;  sleep,  472 
in  acromegaly,  479;  arthritis,  484;  blood  diseases,  492; 
cancer,  503;  chlorosis  and  anaemia,  492;  diabetes 
mellitus,  510;   endoarteriitis,  512;     epilepsy,  541; 
gout,  522;  icterus,  527;  insanity,  543;  leukaemia, 
493;     mental    and  other  nervous     disorders,    536; 
mental  and  other  nervous  disorders  influenced  by 
administration  of  phosphorus  compounds  and  other 
drugs,  545;  morbus  based owii,  581;  nephritis,  547; 
paralysis,  541;     pneumonia,     515;     rachitis,  566; 
rheumatism,  484;  tuberculosis,  583 
See  also  Phosphorus  balances  and  Phosphorus  require- 
ments 
of  foods,  76 

lipoids,  constancy  of,  in  cells  and  organs,  281 
milk,  153,  381 

as  affected  by  foods,  379 
silage  corn  with  and  without  added  floats,  and  of  silage  made 

from  the  same,  102 
the  blood,  149 

changes  in  disease,  496 
bones  as  influenced  by  acidified  hay,  387 

alkaline  earth     carbonates     and  sul- 
phate added  to  oat  rations,  390 
diets  of  hay,  hay  and  oats,  oats  or 

oats  plus  NaH2P04,  389 
lime-  and  phosphorus-poor  diet  with 
and    without    added   phosphates, 
386 
lime-poor  diet,  387 
teeth,  marrow  and  cartilage,  111 
brain  as  affected  by  arteriosclerosis,  135 

chronic  tuberculosis,  588 
nerves  and  cerebrospinal  fluid,  127 
diet,  advantage  of  complexity  of  organization,  355 
digestive  secretions,  172 
feces,  194 

of  infants,  conditions  affecting,  459 
See  also  Phosphorus  Balances 
genitalia,  146 
hypophysis,  146 
kidney,  141 

as  influenced  by  chronic  tuberculosis,  588 
liver,  136 

as  influenced  by  chronic  tuberculosis,  588 
muscles,  120 
spleen,  139 

suprarenal  capsules,  143 
thymus,  143 
thyroid,  144 


INDEX  741 

Phosphorus  of  urine  as  gauge  of  digestive  disturbances  in  infants,  564,  565 
*      F  influenced  by  the  crisis  of  disease,  499 

in  arthritis  and  chronic  rheumatism,  484 
minimum  amount  inferred  from  amniotic  fluid  content,  457 
organic,  in  urine  after  epileptic  attack,  541 

w  as  affected  by  degenerative  nervous  diseases,  5d9 

a  pathological     symptom     in  the     breast-fed 
infant,  565 
output  in  urine,  as  influenced  by  age  of  subject,  407 

artificial  dyspnoea,  479 

exercise,  464 

extirpation  of  pancreas,  511 

fast,  399,  439,  440,  444 

fever,  514 

mental  work  and  by  diet,  461 

rest  and  exercise,  461 
sleep,  472 

thirst  and  water-drinking,  473 
time  of  day,  474 
of  breast-fed  infants,  456,  457 

horse,  422 
See  also  Phosphorus  Balances 
requirement  as  judged  by  excretion  during  fast,  399 
of  animals,  general  discussion,  397 

cattle,  419;  dogs,  426;  horses,  422;  infants,  414;  adult 
human  beings  under     normal     conditions,  403; 
men,  487,  512;  rats,  428;  sheep,  425;  swine,  424 
See  also  Phosphorus  Balances 
retention,  agents  for  reducing,  480 

as  affected  by  dyspepsia,  459 
during  pregnancy,  472 

underfeeding  and  overfeeding,  433 
in  relation  to  typhoid  convalescence,  434 
starvation,  effects  on  tissues  of  the  body  and  on  phosphorus  excre- 
tion, 436 
with  dogs,  394 

milch  cows,  394 
rats,  430 
therapy   in    diseases  of  the  blood,  501 
epilepsy,  547 

malnutrition  of  the  bones,  535 
mental  and  other  nervous  diseases,  545 
osteomalacia,  558,  559 
rachitis,  567 
tuberculosis,  585 
Phytase,  definition,  57 

occurrence,  185,  310 
Phytate  of  calcium,  feeding  experiment,  314,  324 

sodium,  toxic  effects,  314 
Phytic  acid  formula,  53 

synthesis,  54 
Phytin,  chemistry,  51 

cleavage  by  enzyme,  57,  185 
constitution,  54 
clinical  experiments,  315 
effects  in  polyneuritis,  486 

on  composition  of  parts  of  the  body,  327,  370 
character  of  bones,  328 
digestion,  314 
diuresis,  307 

excretion  of  calcium,  magnesium  and  phosphorus,  206 
glycogen  of  the  liver,  305 


742  INDEX 

Phytin,  effects  on  hydrochloric  acid  of  the  stomach,  551 
leucocytosis,  533 
milk,  composition,  381 

and  flow,  307,  312,  315,  383 
nitrogen  and  phosphorus  metabolism,  487,  311 
paths  of  elimination  of  calcium  and  magnesium,  207 
power  of  oxidation,  280 
feeding  experiments,  305,  326,  328,  343-354,  486 
in  bones,  82;  cereal  products,  79,  83,  84 

feces,  196;  seeds,  79,  80,  81,  82 
influence  on  phosphaturia,  315 

under  antirabes  treatment,  566 
occurrence,  51,  81 
of  corn,  chemistry,  53 

cottonseed  meal,  chemistry,  53 
foods,  81 

oats,  chemistry,  53 
laxative  effects,  310,  312 
metabolism,  305 
pharmacodynamic  value,  316 
therapy,  effect  on  erythrocytes  and  haemoglobin,  533 

in  rachitis,  569 
utilization,  influenced  by  species,  197 
Phyto-vitellins,  investigations,  48 
Pituitary  gland.     See  Hypophysis 
Plant  phosphatids,  chemistry,  73 

Plants,  distribution  of  phosphorus  compounds,  84,  87 
Plasmon,  description,  92 

feeding  experiments,  239,  274,  290,  550 
with  phosphorized  cod-liver  oil  for  rachitis,  576 
Pneumonia,  croupous,  effect  on  phosphorus  of  the  urine,  193 

effects  on  composition  of  urine,  organs  and  blood,  499,  500,  507,  515, 
517,  519 
Polycythaemia  rubra  megalosplenica,  lecithin  of  the  blood,  500 
Polyneuritis,  cause,  effects  and  cure,  486 
Polynucleic  acids,  composition,  23 
definition,  22 
structure,  25 
Polynucleotides,  definition,  22 
Poppy  seed  cake,  nuclein  phosphorus,  90 
Potassium  iodide  treatment  in  acromegaly,  increased  calcium  and  phosphorus 

elimination,  482 
Potato,  sweet,  mineral  analyses,  78 

white,  mineral  analyses,  78 
Precipitated  calcium  phosphate,  effect  on  character  of  bones,  328 
Pregnancy,  diet  of  the  mother  not  reflected  in  the  composition  of  the  young,  471 
effect  on  phospholipins  of  ovary  and  corpus  luteum,  148 

phosphorus  requirement,  389 
lipoid  content  of  the  blood  of  the  mother  and  child,  472 
nitrogen  and  phosphorus  ratio  in  intake,  urine  and  feces,  472 
phosphorus  metabolism  during,  471 
storage  during,  408 
Prophylactic  use  of  nucleic  acids,  258 
Proprietary  preparations  of  phosphorus  compounds,  descriptions,  92 

physiological  effects,  550 
Protagon,  chemistry,  72 
Protein-free  milk,  characteristic  effects,  359,  360 

preparation,  359 
Proton,  feeding  experiments,  322,  407 
rectal  feeding  experiment,  404 
Protylin,  description,  93 
effects,  301,  551 


INDEX  743 

Protylin,  feeding  experiments,  301,  324 
preparation  and  properties,  551 
use  in  anaemia,  301 

rachitis,  301,  569 
scrofula,  301 
Prunes,  mineral  analyses,  78 
Pseudonucleins,  definition,  14 

from  phosphoproteins,  33 
Pseudorachitis  distinguished  from  true  rachitis,  513 

effect  on  phosphorus  and  calcium  metabolism,  570,  575 
Psychasthenia,  phosphate  therapy,  545 
Psychatoxies,  phosphate  therapy,  545 
Psychoses,  treatment  with  inorganic  salts,  545 
Purin  bases,  composition,  17 

excretion  after  nucleic  acids,  240,  242,  245 

during  and  after  muscular  activity,  469 
on  light  and  dark  meat,  525 
metabolism.    See  also  Gout 
Purins  of  the  feces,  due  to  bacteria,  234 

urine,  origin,  246 
Pyrimidin  bases,  composition,  19 

nucleotides,  digestion,  233,  236 
Pyro-  ortho-  and  meta-phosphoric  acids  compared  as  to  poisonous  nature,  221 
Pyrophosphates,  poisoning  from,  226,  227 

elimination  of,  221 
Pyrophosphoric  acid  in  vegetable  substances,  91 

poisoning  from,  221,  227 
Rabbit,  organic  phosphorus  compounds,  107 
Rachitis  and  calcium  metabolism,  567 

distinguished  from  malnutrition  of  the  bones,  573 
etiology,  567 

as  phosphorus  metabolism  disturbance,  485 
phosphorus  metabolism  and  phosphorus  treatment,  567 
treatment,  567 

lecithin,  531 
protylin,  301 
Rape  cake,  lecithin  phosphorus,  82,  91 
nuclein  phosphorus,  90,  91 
organic  phosphorus,  82 
phosphorus,  82,  91 
seeds,  inorganic  phosphorus,  79 
lecithin  phosphorus,  79,  82 
organic  phosphorus,  82 
phosphorus,  79 
protein  phosphorus,  79 
Reaction  of  body  tissues  and  fluids,  relations  of  phosphates  to,  178 
Red  dog  flour,  mineral  analyses,  77 
Rennet  coagulation  of  casein,  Bang's  interpretation,  40 

Bosworth's  interpretation,  40 
conditions  favorable,  39 
Hammarsten's  interpretation,  39 
in  the  stomach,  261 
Mellanby's  interpretation,  41 
Schryver's  interpretation,  41 
Rennin.     See  Chymosin 
Rheumatism,  chronic,  forms  of  phosphorus  in  urine,  484 

phosphoric  acid  treatment,  224,  484 
Rice  bran,  phosphorus  content  and  distribution,  82 
diet,  phosphorus  balances,  486,  488 
flour,  phosphorus  content  and  distribution,  79 
mineral  analyses,  77 


744  INDEX 

Rice,  phosphorus  content  and  distribution,  80,  82,  491 

as  affected  by  milling  processes,  78,  79,  488 
polish  and  wheat  bran,  specific  effects  on  swine,  376 
(polished)  diet  as  cause  of  beriberi,  485 
straw,  ether  extract  phosphorus,  90 
Roborat,  description,  93- 

feeding  experiments,  550 
Rock  phosphate,  effects  on  growth,  227 

metabolism  and  slaughter  experiments,  328 
Roughage  compared  with  cereals  and  mill  feeds,  373 
Rye  grains,  inorganic  phosphorus,  79 
lecithin  content,  87 

(phosphatid)  content,  85 
phosphorus,  79 
phosphorus,  79,  87 
protein  phosphorus,  79 
Sahidin,  composition,  69 
Saliva,  composition,  175 

Salicylic  acid,  phosphorus  retention  after,  368 

Salts  of  sodium  and  potassium,  effects  on  phosphorus  excretion,  200 
Sanatogen,  composition,  92 

feeding  experiments,  292,  325 
therapy,  532 
Sanose,  composition,  92,  273 

feeding  experiments,  273 
Sciatica,  glycerophosphate  treatment,  521 

Sclerosis  multiplex,  phosphorus  content  of  cerebrospinal  fluid,  537 
Scrofula,  nuclein  therapy,  257 
protylin  therapy,  301 
Secretion  of  phosphorus  into  alimentary  tract,  182 
Seeds,  distribution  of  phosphorus  compounds,  77,  83,  84 

See  also  the  individual  seeds  and  grains 
Sesame  cake,  lecithin  (phosphatid)  content,  85 

seeds,  phosphorus  content  and  distribution,  82 
Sheep,  mineral  analyses  of  entire  body,  106 
Silage,  addition  of  phosphates,  101 
Skimmed  milk  added  to  corn,  specific  effects  on  swine,  376 

and  cream,  compared  with  whole  milk  with  regard  to  phospho- 
rus compounds,  163 
mineral  analyses,  78 
Sleep,  influence  on  phosphorus  metabolism,  432,  472 
Sorghum,  ether  extract  phosphorus,  90 
Soy  bean  cake,  phosphorus  content  and  distribution,  91 
hay,  mineral  analyses,  78 
beans  added  to  corn,  specific  effects  on  swine,  376 
mineral  analyses,  78 
phosphorus  and  lecithin  content,  87 

(phosphatid)  content,  85 
Species,  effects  on  the  paths  of  phosphorus  excretion,  211 
Sperm,  fish,  phosphorus  content  and  distribution,  148 
Spermatozoa,  phosphorus  compounds,  148 
Sphingomyelin,  chemistry,  70 

in  blood  cells,  occurrence,  153 
Spleen,  ash  analyses,  105 

composition  as  affected  by  various  diseases,  507 
lecithin  content,  107,  140 
mineral  analyses,  507 
nuclein  phosphorus,  140 
nucleon  content,  140 
nucleoprotein,  140 

phosphatid  content  as  influenced  by  alcoholism,  484 
■    phosphatids,  588 


INDEX  745 

Spleen,  phosphorus,  139, 140, 149,  507,  588 

phosphorus  content  and  distribution  in  lipoids  as  influenced  by  chronic 

tuberculosis,  588 
partition,  139 

as  influenced  by  age,  139 
removal,  effect  on  metabolism  and  composition  of  body,  580 
Split-protein  feeding  for  infants,  455 
Springer's  cereal  decoction,  description,  92 
Sterilized  and  raw  milk,  compared,  in  vitro  digestions,  274 

metabolism  experiments,  415 
milk,  infant  feeding  experiments,  414,  453,  454 
Stored  protein,  Kolpakcha's  idea,  216 

Striated  and  non-striated  muscles,  comparisons  of  phosphorus  content,  123 
Strontium,  substitution  for  calcium  in  bone,  386,  390 

treatment  in  rachitis,  568 
Sulphur  elimination  as  related  to  nitrogen  and  phosphorus  elimination,  220 
Sunflower  seeds,  phosphorus  and  phosphatid  content,  81,  85 
Superphosphate,  feeding  experiments,  386 

Suprarenal  capsules,  phosphorus  content  and  partition,  107,  143 
Sweating,  influence  on  phosphorus  excretion,  466,  467 
Sweet-breads,  feeding  experiments,  324 
Swine,  mineral  analyses  of  entire  body,  106 
Synthesis  of  glycerophosphates,  74 

lecithin  in  developing  eggs,  341 

egg  production,  342 
nucleins  as  leucocytes,  229 
disussion,  30 
enzymes  effecting,  230 
in  developing  eggs,  229,  341 
fish  genitalia,  229 
suckling  animals,  229 
on  purin-free  diet,  229 
organic  phosphorus  compounds  in  fasting  fish,  354 

from  inorganic,  animal  investiga- 
tions, 318 
discussion  and  animal 
experiments,  318 
phosphate-carbohydrate  compounds,  31 

-protein  compounds,  30 
phosphatids,  74 

in  brain  development,  341 
phytic  acid,  54 
Synthetic  glycerophosphoric  acid,  absorption,  293 
Szekely's  milk,  composition,  454 

infant  feeding  experiments,  454 
Tankage,  mineral  analyses,  78 
Teeth,  composition,  115 

as  influenced  by  caries,  504 
fast,  441 
dentine,  mineral  analyses,  113,  116 

phosphorus,  113,  116  .    ■ 

enamel,  mineral  analyses,  113,  116,  117 

phosphorus,  113,  116,  117 
influenced  by  hard  water,  394 

lime-poor  diet  less  than  bones,  388 
mineral  analyses,  113,  313,  505 
organic  phosphorus,  115 
phosphorus,  113,  313,  505 

relation  of  parathyroids  to  dentine  formation,  560 
Tetra-ethyl-phosphonium  iodide,  injection  experiments,  549 
Therapeutic  and  prophylactic  use  of  nucleins  and  nucleic  acids,  256,  502,  504, 
519,  547,  586 
use  of  phytin,  315,  533,  569 


746  INDEX 

Thudichum's  classification  of  the  brain  constituents,  128 
Thymic  acid,  definition,  20 
Thymin,  composition,  19 
Thymonucleict  acids,  definition,  19 
Thymus,  feeding  experiments,  324 

nucleic  acid,  structure,  26,  27 
phosphorus  content  and  partition,  143,  149 
Thyraden  treatment  for  osteomalacia,  558 
Thyrein.     See  Iodothyrin 
Thyreiodin.     See  Iodothyrin 
Thyreoglobulin,  occurrence  and  significance,  145 
Thyreoidin.     See  Thyroidin,  545 
Thyroiodinin.     See  Iodothyrin 

Thyreonucleoalbumin,  occurrence  and  significance,  144 
Thyroid  glands  and  hypophysis,  reciprocal  functions,  481 
in  relation  to  phosphorus  metabolism,  577 
investigations  as  to  the  active  component,  145 
preparations,  descriptions  and  names,  545 
treatment,  influence  on  bone  formation,  559 

physiological  effects,  577 
treatment  in  acromegaly,  479 

influence  on  phosphorus  and  calcium  excre- 
tion, 480,  482 
cretinism,  546 
exophthalmic  goiter,  581 
leukaemia,  493 
mongoloid  idiocy,  546 
myxoedema,  546 
See  also  Iodothyrin,  Thyraden  and  Thyroidin 
Thyroids,  effects  on  general  metabolism,  577 
phosphorus  and  iodine  content,  579 

content  and  partition,  144,  149 
Thyroidectomy,  effects  on  nuclease  content  of  organs  and  blood,  235 
bone  formation,  559 
physiological  effects,  577 
Thyroidin,  distinction  from  iodothyrin,  545 

treatment  after  thyroidectomy,  580 

effect  on  phosphorus  excretion  by  a  thyro-parathyroid- 

ectomized  dog,  579 
in  paralysis  agitans,  545 

senility  and  paralysis  agitans,  phosphorus  balances,  545 
Thyroiodin  distinguished  from  thyroidin,  545 
Thyroiodinin.     See  Iodothyrin 
Thyro-parathyroidectomy,  effect  on  partition  of  phosphorus  of  the  blood,  560 

phosphorus  elimination,  561 
Time  of  day,  effect  on  phosphorus  metabolism,  474 
Timothy  hay,  mineral  analyses,  78 
Torulin  as  cure  for  beriberi,  490 

composition,  490 
Triamino-diphosphatids,  chemistry,  69 

-monophosphatids,  chemistry,  71 
Tricalcol,  absorption,  184 
Triferrin,  effects  on  composition  of  liver,  139 
Triticonucleate  of  sodium  treatment  in  tuberculosis,  587 
Triticonucleic  acid,  chemistry,  91 
effects,  250 
in  wheat  germ,  91 
treatment  in  tuberculosis,  257 
Tropon,  feeding  experiments  with  a  man  and  a  dog,  550 
Tuberculin  treatment  in  tuberculosis,  584 

Tuberculosis  and  chlorosis,  phosphorus  and  chlorine  elimination,  584 
chronic,  changes  in  lipoids  of  organs,  587 
demineralization,  584 


INDEX  747 

Tuberculosis,  effect  on  lipoids  and  phosphorus  of  organs — guinea  pigs,  588 
phosphorus  of  milk,  382 
lecithin  therapy,  530,  531,  532 
nucleic  acid  and  disodium-methyl-arsenate  therapy,  586 

therapy,  257 
nuclein-saline  treatment,  586 
phosphaturia  in.     See  Phosphaturia 
phosphorus  metabolism  and  phosphorus  treatment,  583 
with  melancholia,  phosphorus  content  of  cerebrospinal  fluid,  537 
Tumor  on  the  brain,  phosphorus  content  of  cerebrospinal  fluid,  537 
Turkish  baths,  influence  on  phosphate,  uric  acid,  urea,  sulphate  and  chloride 

excretion,  466 
Turnips,  phosphorus  content  and  partition,  88 

as  affected  by  fertilizer,  100 
Tryptic  digestion  of  casein,  influence  of  alkalinity,  272 

in  relation  to  ionproteid  theory,  271 
in  vitro  tests,  267 
nucleoproteins,  231 
ovovitellin,  271 
Typhoid  fever,  effects  on  composition  of  blood,  500 

and  urine,  499 
Ulcer  of  the  stomach,  phytin  ingestion,  316 
Uracil,  composition,  19 
Uric  acid,  composition,  18 

elimination  after  nucleic  acid  ingestion,  245 
See  also  Gout 
Urinary  elimination  of  phosphorus,  method,  188 
Urine,  composition  as  affected  by  vegetable  diet — several  species,  211 

See  also  Phosphorus  balances,  and  many  feeding  experiments 
excessive  phosphorus.     See  Phosphaturia 

glycerophosphoric  acid  content  in  fatty  degeneration  explained,  513 
organic  phosphorus  content,  190 

as  affected  by  croupous  pneumonia,  193 

degenerative  nervous  diseases, 

539 
epileptic  attack,  541 
a  pathological  symptom  in  the  breast-fed 
infant,  565 
of  infant's,  456 
phosphoric  acid  esters,  191 
phosphorus  content  as  affected  by  calcium  salts,  207 

a  gauge  of  digestive  disturbances  in  infants,  564, 
565 
ratio  of  phosphorus  to  nitrogen.     See  Nitrogen  and  phosphorus  ratio  in 

urine 
Vegetable  casein,  feeding  experiment,  322 

diet,  effects  on  phosphorus  elimination,  208 

human  urinary  nitrogen  and  phosphorus,  209,  210 

as  influenced  by  mental 
work,  461 
phosphorus  and    alkali    phosphates    as 
T,  ,  ,   ,  •  influenced  by  physical  exercise,  465 

Vetch  hay,  ether  extract  phosphorus,  90 
Vitamine,  definition,  365 
Vitellin,  cleavage  products,  46 
in  eggs,  448 

feeding  experiments,  320,  321,  322 
loss  in  incubating  eggs,  447 
Water-drinking  and  thirst,  influence  on  phosphorus  metabolism,  473 

effects  on  composition  of  forage  plants,  93 
Water  in  the  organs  in  disease,  509 

on  the  brain,  phosphorus  content  of  cerebrospinal  fluid,  537 


748  INDEX 

Wheat  bran  and  rice  polish,  specific  effects  on  swine,  376 
mineral  analyses,  77 

phosphorus  content  and  partition,  77,  82,  85 
germ  flour,  phosphorus  content  and  distribution,  78 
lecithin  (phosphatid)  content,  85 
mineral  analyses,  77 
tritico-nucleic  acid,  91 
gluten,  mineral  analyses,  77 
grains,  inorganic  phosphorus,  77,  79 

as  affected  by  fertilizer,  100 
lecithin  content,  87 

(phosphatid)  content,  85 
phosphorus,  79,  82 
mineral  analyses,  77 

as  affected  by  fertilizer,  100 
organic  phosphorus,  82 
phosphorus,  77,  79,  87,  99 

as  affected  by  fertilizer,  97,  100 
protein  phosphorus,  79 
inadequacy  for  nutrition,  384,  385 
middlings  added  to  corn,  specific  effects  on  swine,  376 

mineral  analyses,  77 
phosphorus,  references,  76 
straw,  mineral  analyses,  78 

phosphorus  as  affected  by  fertilizer,  96 
Whey,  in  infant  feeding,  458 
mineral  analyses,  78 
modification  of  milk;  effect  on  mineral  retention,  459 

phosphorus  elimination,  459 
protein,  definition,  39 
Whole  milk,  skimmed  milk  and  cream,  phosphorus  compounds,  163 
Wine,  effect  on  nitrogen  and  phosphorus  elimination,  469 
Xanthin  bases.     See  Purin  bases 

'  composition,  18 
Xanthooxidase,  definition,  28 
Yeast  cures  polyneuritis,  489 
endotrypsin,  234 
nucleic  acid,  structure,  26 
Zein,  maintenance  and  slaughter  experiments,  331 


COLUMBIA  UNIVERSITY  LIBRARIES 

This  book  is  due  on  the  date  indicated  below,  or  at  the 
expiration  of  a  definite  period  after  the  date  of  borrowing, 
as  provided  by  the  rules  of  the  Library  or  by  special  ar- 
rangement with  the  Librarian  in  charge. 



DATE   BORROWED 

DATE  DUE 

DATE  BORROWED 

DATE   DUE 

1955 

C2BI1  140IM100 

j 

QP535.P1  F74 

Forbes 

A  review  of  the  literature  of 


